Composition for inducing proliferation or accumulation of regulatory t cells

Technical field] [0001] The document describes a composition that affects the induction of regulatory T cell proliferation or accumulation and which comprises, as an active ingredient, bacteria belonging to the genus Clostridium, a physiologically active substance derived from bacteria, sporic forms of bacteria and the like. The invention relates to a composition for use in a method of treating or preventing a disease selected from an infectious disease, autoimmune disease or allergic disease in a subject, a composition comprising as active ingredient bacteria, including spore-forming bacteria belonging to clusters IV and XIVa of Clostridium in combination, which the combination induces the proliferation or accumulation of the Foxp3 transcription factor positive regulatory T lymphocytes in the subject.

[Background of the invention] [0002] Hundreds of species of commensal microorganisms thrive in the gastrointestinal tract of mammals, closely interacting with the host immune system. The results of studies using microbial-free animals (GF) have shown that commensal microorganisms exert enormous influence on the development of mucosal immune systems, such as histogenesis of Peyer's patches (PPS) and isolated lymphoid vesicles (ILFs), secretion of antimicrobial peptides from epithelium and accumulation of specific lymphocytes in mucosal tissues, specific lymphocytes including immunoglobulin A produced by plasmocytes, intraepithelial lymphocytes, IL-17 produced by CD4 positive T cells (Th17) and IL-22 produced by cells such as NK (non-patent documents 1 to 7). Respectively, the presence of intestinal bacteria increases the protective effect of mucous membranes, providing the host with strong immune responses against the invasion of pathogenic microorganisms. On the other hand, mucosal immune systems maintain a lack of response to food antigens and harmless microorganisms (non-patent document 3). For this reason, irregularities in the regulation of mutual communication between commensal bacteria and the immune system (intestinal dysbiosis) can cause too strong immune response to environmental antigens, leading to inflammatory bowel disease (IBD) (non-patent documents 8 to 10). mucosal immune systems maintain a lack of response to food antigens and harmless microorganisms (non-patent document 3). For this reason, irregularities in the regulation of mutual communication between commensal bacteria and the immune system (intestinal dysbiosis) can cause too strong immune response to environmental antigens, leading to inflammatory bowel disease (IBD) (non-patent documents 8 to 10). mucosal immune systems maintain a lack of response to food antigens and harmless microorganisms (non-patent document 3). For this reason, irregularities in the regulation of mutual communication between commensal bacteria and the immune system (intestinal dysbiosis) can cause too strong immune response to environmental antigens, leading to inflammatory bowel disease (IBD) (non-patent documents 8 to 10).

[0003] The results of recent studies have shown that individual commensal bacteria control the differentiation of specific immune cells in the mucosal immune system. For example, Bacteroides fragilis, which is a commensal bacterium in humans, especially induces a systemic response of Th1 cells and IL-10 mucosa, triggering a T cell response in mice and protects the host against colitis that would otherwise be caused by a pathogen (Non-Patent Document 3 ). It has been shown that segmented filamentous bacteria, which are intestinal commensal bacteria in mice, induce a mucosal Th17 cell response and thus increase immunity to mice.

- pathogen infections of the host's gastrointestinal tract (non-patent documents 11 to 13). In addition, short-chain fatty acids derived from several commensal bacteria inhibit intestinal inflammation (non-patent document 14). In addition, the presence of certain species of intestinal microbiota is expected to have a major impact on the differentiation of regulatory T lymphocytes (hereinafter referred to as "Treg lymphocytes") that maintain homeostasis of the immune system.

[0004] Meanwhile, regulatory T cells that have been identified as a subset that weakens immunity are CD4 T-lymphocytes⁺that express the Foxp3 transcription factor and that play an important role in maintaining immune homeostasis (non-patent documents 8, 9, 15 and 16). In addition, it is known that the cells expressing Foxp3 are present in large numbers especially in the large intestine, and only the Treg lymphocytes locally in the large intestine permanently express IL-10, which is a high-level immunosuppressive cytokine (Non-patent document 17). It is also known that animals having CD4 cells⁺ Foxp3⁺which especially IL-10 has been removed, develop inflammatory bowel disease (non-patent document 18).

[0005] Accordingly, if the mechanism of induction of Treg lymphocytes that produce IL-10 in a large amount in the large intestine is explained, immunosuppression can be enhanced, which, in turn, can be used to treat autoimmune diseases, such as inflammatory bowel disease. and in organ transplantation.

[0006] However, mechanisms affecting how much of the Treg lymphocyte is present in the large intestine and what is the mechanism by which the Treg lymphocytes produce IL-10 in large quantities in the large intestine are still unclear. In addition, it is still unclear which bacterial species that make up the commensal microbiota bacteria influence the induction of regulatory T-lymphocytes.

[Quoted sources] [Non-patent literature] [NPL 1] JJ Cebra, "Am J Clin Nutr", May, 1999, 69, 1046S.

[NPL 2] AJ Macpherson, NL Harris, "Nat Rev Immunol", June 2004, 4, 478.

[NPL 3] JL Round, SK Mazmanian, "Nat Rev Immunol", May 2009, 9, 313.

[NPL 4] D. Bouskra et al., "Nature", November 27, 2008, 456, 507.

[NPL 5] K. Atarashi et al., "Nature", October 9, 2008, 455, 808.

[NPL 6] Ivanov, II et al., "Cell Host Microbe", October 16, 2008, 4, 337.

[NPL 7] SL Sanos et al., "Nat Immunol", January 2009, 10, 83.

[NPL 8] MA Curotto de Lafaille, JJ Lafaille, "Immunity", May 2009, 30, 626.

[NPL 9] MJ Barnes, F. Powrie, "Immunity", September 18, 2009, 31, 401.

-3 [NPL 10] WS Garrett et al., "Cell", October 5, 2007, 131, 33.

[NPL 11] Ivanov, II et al., "Cell", October 30, 2009, 139, 485.

[NPL 12] V. Gaboriau-Routhiau et al., "Immunity", October 16, 2009, 31, 677.

[NPL 13] NH Salzman et al., "Nat Immunol", 11, 76.

[NPL 14] KM Maslowski et al., "Nature", October 29, 2009, 461, 1282.

[NPL 15] LF Lu, A. Rudensky, "Genes Dev", June 1, 2009, 23, 1270.

[NPL 16] S. Sakaguchi, T. Yamaguchi, T. Nomura, M. Ono, "Cell", May 30, 2008, 133, 775.

[NPL 17] CL Maynard et al., "Nat Immunol", September 2007, 8, 931.

[NPL 18] YP Rubtsov et al., "Immunity", April 2008, 28, 546.

[0008] V. Gaboriau-Routhiau et al., "Immunity", October 16, 2009, 31, 677 describes segmented filamentous bacteria (SFB) that do not belong to the genus Clostridium, and which are actually phylogenetically distant from them.

[0009], Sokol Harry et al: "Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patients", Proceedings of the National Academy of Sciences, National Academy of Sciences, Washington, DC; U. S., vol. 105, no. 43.28 October 2008 (2008-10-28), pages 16731-16736, he proposes to Faecalibacterium prausnitzii (belonging to Clostridium cluster IV) as a candidate for a probiotic agent in the treatment of Crohn's disease (CD).

[0010] DE 10 2006 062250 A1 (Saur-Brosch Roland [DE]) discloses the use of a composition comprising minerals and / or vitamins and, optionally, acetogenic and / or butyrogenic bacteria for oral or rectal administration in the treatment or prevention of abdominal discomforts, which are associated with reduced reduction acetic metabolic activity.

[0011] Itoh K et al: "Characterization of clostridia isolated from the flora of the limited mica and their effect on caecal size when associated with germ-free mice". Laboratory Animals KWI 1985 vol. 19, no. 2, April 1985 (1985-04), pages 111-118 discloses the characteristics of 115 Clostridium strains accumulated from 3 separate stool samples and reduced microbiota (LF) of mice cultured by inoculation into mice free from faecal microbes of conventional mice that have been exposed to chloroform and their effect on the size of the cecum in microbial-free mice.

[Summary of the Invention] [Technical Problem] [Solution to the Problem] [0012] The inventors have found that the fraction treated with chloroform and the fraction forming spore forms in a mammalian stool sample induce accumulation of regulatory T lymphocytes (Treg lymphocytes) in the large intestine. In addition, the inventors found that the bacteria

- belonging to the genus Clostridium induce the proliferation or accumulation of regulatory T cells in the large intestine. The inventors have also found that regulator T lymphocytes induced by these bacteria attenuate the proliferation of effector T cells. In addition, the inventors also found that colonization of bacteria belonging to the genus Clostridium and resulting in the proliferation or accumulation of Treg lymphocytes, regulates local and systemic immune responses.

[0013] Based on the findings, the inventors have found that the use of bacteria belonging to the genus Clostridium, their spore forms or a physiologically active derivative allows induction of regulatory T cell proliferation or accumulation (Treg lymphocytes) and further weakening of immune function.

[0014] In one aspect, the invention provides a composition for use in a method of treating or preventing a disease selected from an infectious disease, autoimmune disease or allergic disease in a subject, wherein the composition comprises, as an active ingredient, spore-forming bacteria belonging to clusters IV and XIVa Clostridium in combination which induces the proliferation or accumulation of the transcription factor Foxp3 positive regulatory T lymphocytes in a subject.

[0015] Other aspects of the invention are defined in the appended claims.

[Preferred effects] [0016] The compositions may serve as an excellent composition for inducing the proliferation or accumulation of regulatory T cells (Treg cells). Resistance in a living body can be reduced by administering the composition as a pharmaceutical product or by ingesting the composition in the form of a food or beverage. Accordingly, the composition can be used, for example, for the prevention or treatment of autoimmune diseases or allergic diseases, as well as for the prevention of immune rejections in organ transplants and the like. Additionally, in the case where the food or beverage, such as health food, contains a composition of the invention, healthy persons can easily and routinely take the composition. Consequently,

[Brief Description of the Figures] [Fig. 1] Fig. 1 is a schematic diagram showing the culture of Il10 mice^venus.

[Fig. [Fig. 2] Fig. 2 is a diagram showing the results of Southern blotting performed to analyze whether Il10 mice^venus they have the Il10 allele^venus.

[Fig. 3] Fig. 3 is a FACS scatterplot showing results obtained when sorting Venus positive cells and Venus negative Il10 mice^venus.

-5 [Fig. 4] Fig. 4 is a graph showing the results obtained when the amounts of IL-10 mRNA expressed on Venus positive cells and Venus negative Il10 mice^venus were analyzed using RT-PCR in real time.

[Fig. 5] Fig. 5 is a graph showing change in the ratio of Foxp3 cells⁺ in CD4 lymphocytes⁺ in SPF mice.

[Fig. [Fig. 6] Fig. 6 shows FACS point diagrams showing the results of the analysis of the Foxp3 cell quantitative ratio⁺ in CD4 lymphocytes⁺ isolated from the small intestine, large intestine and peripheral lymph nodes of GF mice and SPF mice.

[Fig. [Fig. 7] Fig. 7 is a graph showing the results of the analysis of the ratio of Foxp3 cells⁺ in CD4 lymphocytes⁺ isolated from the small intestine, large intestine and peripheral lymph nodes of GF mice and SPF mice.

[Fig. [Fig. 8] Fig. 8 is a graph showing the results of the analysis of the number of CD4 cells⁺ Foxp3⁺isolated from the small intestine, large intestine and peripheral lymph nodes of GF mice and SPF mice.

[Fig. [Fig. 9] Fig. 9 is a line graph showing the results of the analysis of the Venus cell quantitative ratio⁺ in CD4 cells⁺ in various tissues of SPF mice treated with antibiotics.

[Fig. [Fig. 10] Fig. 10 shows FACS point diagrams showing the results of the ratio analysis of Foxp3 cells⁺ in CD4 lymphocytes⁺ isolated from the lamina propria of the GF mouse, where the SPF mouse stool suspension was administered.

[Fig. 11] Fig. 11 is a graph showing the results of the analysis of the ratio of Foxp3 cells⁺ in CD4 lymphocytes⁺ isolated from the lamina propria of the large intestine and the lamina propria of the small intestine of the GF mice receiving the SPF mouse faecal suspension.

[Fig. [Fig. 12] Fig. 12 is a graph showing the results of the analysis of the ratio of Foxp3 cells⁺ in CD4 lymphocytes⁺ isolated from the lamina propria mice deficient in ILF, PP and tufts of the large intestine.

[Fig. [Fig. 13] Fig. 13 shows FACS point diagrams showing the results of the analysis of the Foxp3 cell quantitative ratio⁺ in CD4 lymphocytes⁺ isolated from the lamina propria of GF mice, given specific commensal bacteria.

[Fig. [Fig. 14] Fig. 14 is a graph showing the results of a ratio analysis of Foxp3 cells⁺ in CD4 lymphocytes⁺ isolated from the lamina propria of GF mice, given specific commensal bacteria.

[Fig. [Fig. 15] Fig. 15 is a graph showing the results of an analysis of the quantitative ratio of IFN-γ in CD4 lymphocytes⁺ isolated from the lamina propria of the large intestine mice, in which specific commensal bacteria were colonized.

[Fig. [Fig. 16] Fig. 16 is a graph showing the results of an analysis of the quantitative ratio of IL-17 cells⁺ in CD4 lymphocytes⁺ isolated from the lamina propria of large intestine in mice with specific commensal bacteria colonized.

-6 [Fig. 17] Fig. 17 is a graph showing the results of the analysis of the ratio of Foxp3 cells⁺ in CD4 lymphocytes⁺ isolated from the large intestine of SPF mice deficient in factor associated with the recognition of molecular patterns of pathogens.

[Fig. 18] Fig. 18 is a graph showing the results of the analysis of the ratio of Foxp3 cells⁺ in CD4 lymphocytes⁺ isolated from the lamina propria of the Myd88 mouse^{- / -}who were colonized with Clostridium.

[Fig. [Fig. 19] Fig. 19 shows FACS point diagrams showing the results of the analysis of the Venus cell quantitative ratio⁺ in lymphocytes isolated from various mouse tissues

IL10^venus.

[Fig. 20] Fig. 20 is a FACS scatterplot showing the results of β-cell T cell receptor expression analysis on lymphocyte cell surfaces isolated from the lamina propria of Il10 mouse large intestine^venus.

[Fig. [Fig. 21] Fig. 21 shows FACS point diagrams showing the results of expression analysis

IL-17, IL-4 and IFN-γ in lymphocytes isolated from the lamina propria of the Il10 mouse^venus.

[Fig. [Fig. 22] Fig. 22 is a graph showing the results of the analysis of the amount of mRNA for IL-10, CTLA4, Foxp3 and GITR expressed in Foxp3-CD4 cells⁺ spleen, Foxp3 cells⁺ CD4⁺ spleen, Venus cells⁺ gills of the proper colon and Venus cells⁺ the lamina of the proper small intestine.

[Fig. [Fig. 23] Fig. 23 shows FACS point diagrams showing the results of analysis of CD4, Foxp3 and Venus expression in the lamina propria of the small intestine and the lamina propria of the GF Il10 mouse^venus and SP10 Il10 mice^venus.

[Fig. [Fig. 24] Fig. 24 shows FACS point diagrams showing the results of Venusi Foxp3 expression of CD4 cells in various IL10 SPF mouse tissues^venus.

[Fig. [Fig. 25] Fig. 25 shows FACS point diagrams showing the results of Foxp3 and Venus expression analysis in Il10 mice^venusfor which specific commensal bacteria were colonized.

[Fig. 26] Fig. 26 is a graph showing the results of analysis of the expression of Foxp3 and / or Venus of CD4 + cells in the small intestine of Il10 mice^venusfor which specific commensal bacteria were colonized.

[Fig. 27] Fig. 27 is a graph showing the results of analysis of the expression of Foxp3 and / or Venus of CD4 + cells in the large intestine of Il10 mice.^venusfor which specific commensal bacteria were colonized.

[Fig. 28] Fig. 28 is a scatter diagram showing the results of the analysis of the ratio of the Venus + cells in CD4 + cells isolated from various Il10 mouse tissues^venuswho were given antibiotics.

[Fig. 29] Fig. 29 is a graph showing the results of the analysis of immunoregulatory functions of CD4 + Venus + cells of the GF Il10 mouse large intestine^venusin which

A type of Clostridium, CD4 cells was colonized⁺ venus⁺ lamina propria of the large mouse SPF Il10^venus and CD4 + GFP + cells from the spleen of the Foxp3 reference mouse^eGFP [Fig. [Fig. 30] Fig. 30 is a graph showing the results obtained when SP6 B6 mice were dosed with polymyxin B or vancomycin for 4 weeks and then the ratio of Foxp3 cells was analyzed.⁺ in the CD4 cell group⁺.

[Fig. 31] Fig. 31 is a graph showing the results obtained after oral administration to mice

GF of SPF mouse faeces, which was subjected to chloroform and examined the ratio of Foxp3 cells⁺ in the CD4 cell group⁺.

[Fig. [Fig. 32] Fig. 32 is a graph depicting the overall flow cytometry analysis of Helios expression on LP grasic lymphocytes or intestinal thick SPF mice, GF mice, Lactobacillus colonized mice or Clostridium colonized mice.

[Fig. [Fig. 33] Fig. 33 is a schematic diagram showing representative results of flow cytometry analysis of CD4 expression, Foxp3 expression and Helios expression in LP grasic lymphocytes or intestinal SPF mole mice, GF mice, Lactobacillus colonized mice or Clostridium colonized mice.

[Fig. 34] Fig. 34 is a graph showing the results obtained when the whole colon intestine derived from GF mice, colonized Lactobacillus mice or colonized Clostridium mice were sowed and culture supernatants were analyzed for TGF-βΙ concentration by ELISA.

[Fig. [Fig. 35] Fig. 35 is a graph showing the results obtained when the intestinal epithelial cells (IECs) were seeded from GF mice or colonized Clostridium mice, and their culture supernatants were analyzed for TGF ^ 1 concentration by ELISA.

[Fig. [Fig. 36] Fig. 36 is a graph showing the results obtained when CD4 T lymphocytes were seeded⁺ spleen in concert with the anti-CD3 antibody and the IEC culture supernatant isolated in GF mice or mice colonized with 46 bacterial strains of the genus Clostridium (Clost.) in the presence or absence of anti-TGF-β and T-lymphocytes that were harvested on day 5 of culture and expression analyzed Foxp3 using RT-PCR in real time.

[Fig. 37] Fig. 37 is a graph showing the results obtained when C57BL / 6 GF mice were orally inoculated with 46 strains of bacteria of the genus Clostridium (Clost.) Or three strains of bacteria of the genus Lactobacillus (Lacto.), And collected IEC three weeks after inoculation and analyzed relative level of mRNA expression of the MMP2 gene using real-time RT-PCR.

[Fig. 38] Fig. 38 is a graph showing the results obtained when C57BL / 6 GF mice were orally inoculated with 46 strains of bacteria of the genus Clostridium (Clost.) Or three strains of bacteria of the genus Lactobacillus (Lacto.), And collected IEC three weeks after inoculation and analyzed relative level of mRNA expression of the MMP9 gene using real-time RT-PCR.

-8 [Fig. 39] Fig. 39 is a graph showing the results obtained when C57BL / 6 GF mice were orally inoculated with 46 strains of bacteria of the genus Clostridium (Clost.) Or three strains of bacteria of the genus Lactobacillus (Lacto.), And collected IEC three weeks after inoculation and analyzed relative level of mRNA expression of the MMP13 gene using real-time RT-PCR.

[Fig. 40] Fig. 40 is a graph showing the results obtained when C57BL / 6 GF mice were orally inoculated with 46 strains of bacteria of the genus Clostridium (Clost.) Or three strains of bacteria of the genus Lactobacillus (Lacto.), And collected IEC three weeks after inoculation and analyzed the relative expression level of the IDO mRNA using RT-PCR in real time.

[Fig. 41] Fig. 41 is a graph showing results obtained when control mice (SPF) and mice given Clostridium (SPF + Clost.) Were given 2% DSS, weight loss, stool hardness and bleeding were observed and measured over time to six days, and then rated numerically.

[Fig. 42] Fig. 42 is a picture depicting the condition of large intestines assessed on day 6 after administration to control mice (SPF) and mice treated with Clostridium (SPF + Clost.), 2% DSS.

[Fig. 43] Fig. 43 shows micrographs showing results obtained when control mice (SPF) and Clostridium-treated mice (SPF + Clost.) Were given 2% DSS and their intestines collected on day 6 and analyzed by histological staining with hematoxylin and eosin.

[Fig. 44] Fig. 44 is a graph showing results obtained when control mice (SPF) and mice given Clostridium (SPF + Clost.) Were sensitized with oxazolone, followed by 1% oxazolone / 50% ethanol solution recto and weight loss measured .

[Fig. [Fig. 45] Fig. 45 shows micrographs showing results obtained when control mice (SPF) and mice given Clostridium (SPF + Clost.) Were sensitized with oxazolone, followed by 1% oxazolone / 50% ethanol solution rectally, and intestines subjected to them. treatment was analyzed by histological staining with hematoxylin and eosin.

[Fig. 46] Fig. 46 is a graph showing the results obtained when control mice (SPF) and mice given Clostridium (SPF + Clost.) Were immunized with ovalbumin (OVA) twice daily with an interval of 2 weeks, sera and serum collected analyzed for serum concentration of specific IgE OVA by ELISA.

[Fig. 47] Fig. 47 is a graph showing results obtained when control mice (SPF) and mice given Clostridium (SPF + Clost.) Were immunized with ovalbumin (OVA) twice daily with an interval of 2 weeks, spleen cells were harvested and analyzed for IL-4 production by spleen cells by in vitro restimulation of OVA.

-9 [Fig. 48] Fig. 48 is a graph showing results obtained when control mice (SPF) and mice given Clostridium (SPF + Clost.) Were immunized with ovalbumin (OVA) twice daily with an interval of 2 weeks, spleen cells were harvested and analyzed for IL-10 production by spleen cells by in vitro restimulation of OVA.

[Fig. [Fig. 49] Fig. 49 shows a phylogenetic tree obtained by the close neighbor method to obtain sequences of 41 Clostridium strains and known bacteria from the Genbank database using Mega software.

[Fig. [Fig. 50] Fig. 50 is a histogram showing the expression of Foxp3 on GF gated CD4 mouse cells (microbial # 1 and # 2) or GF mice in which three Clostridial strains belonging to cluster IV (3 strains of Clost. Mouse # 1 were colonized; # 2).

[Fig. 51] Fig. 51 is a histogram depicting the expression of Foxp3 through the positive CD4 cells of GF mice (GF) or GF mice, which were administered by gavage treated with human stool treated with chloroform (GF + Chloro.).

[Fig. [Fig. 52] Fig. 52 is a graph depicting the expression of Foxp3 by G4 positive GF (GF) positive mouse lymphocytes or GF mice that were administered by gavage treated with chloroform (GF + Chloro.).

[Fig. [Fig. 53] Fig. 53 is a graph showing the amounts of Clostridium and Bacteroides in the stools of mice that were administered by gavage to a human stool treated with chloroform.

[Description of Embodiments] [0018] "Regulatory T-lymphocytes" refer to the FoxP3 transcription factor positive in CD4 positive T lymphocytes.

[0019] The meaning of "induce the proliferation or accumulation of regulatory T lymphocytes" in the invention includes the effect of inducing differentiation of immature T cells on regulatory T cells, the differentiation of which leads to the proliferation or accumulation of regulatory T lymphocytes. In addition, the meaning "induces the proliferation or accumulation of regulatory T lymphocytes" in the invention includes in vivo effects, in vitro effects, and ex vivo effects. Accordingly, it includes all of the following effects: the effect of inducing the proliferation or accumulation of regulatory T lymphocytes in vivo by administering or ingesting bacteria belonging to the genus Clostridium or a physiologically active substance or a similar bacterial derivative; the effect of inducing the proliferation or accumulation of cultured regulatory T lymphocytes by the action of bacteria belonging to the genus Clostridium or a physiologically active substance or the like, a bacterial derivative, to cultured regulatory T-cells; and the effect of inducing the proliferation or accumulation of regulatory T lymphocytes that have been collected from a living organism and which are then to be introduced into a living organism, such as the organism from which they were obtained or other organism, through the action of bacteria belonging to the genus Clostridium or a physiologically active substance or a similar bacterial derivative, on regulatory T-cells. The effect of inducing the proliferation or accumulation of regulatory T lymphocytes and the effect of inducing the proliferation or accumulation of regulatory T lymphocytes that have been collected from a living organism and which are then to be introduced into a living organism, such as the organism from which they were obtained or other organism, through the action of bacteria belonging to the genus Clostridium or a physiologically active substance or a similar bacterial derivative, on regulatory T-cells. The effect of inducing the proliferation or accumulation of regulatory T lymphocytes and the effect of inducing the proliferation or accumulation of regulatory T lymphocytes that have been collected from a living organism and which are then to be introduced into a living organism, such as the organism from which they were obtained or other organism, through the action of bacteria belonging to the genus Clostridium or a physiologically active substance or a similar bacterial derivative, on regulatory T-cells. The effect of inducing the proliferation or accumulation of regulatory T lymphocytes on regulatory T-cells. The effect of inducing the proliferation or accumulation of regulatory T lymphocytes on regulatory T-cells. The effect of inducing the proliferation or accumulation of regulatory T lymphocytes

You can evaluate, for example, the following. Specifically, bacteria belonging to the genus Clostridium or a physiologically active substance or similar bacterial derivative were orally administered to experimental animals, such as microbial-resistant mice, then CD4-positive cells were isolated in the large intestine and the quantitative ratio of regulatory T-cells contained in positive CD4 cells was measured by cytometry flow (see Example 7).

[0020] Regulatory T-lymphocytes whose proliferation or accumulation is induced by the composition of the invention are Foxp3 transcription factors of positive regulatory T-lymphocytes.

[0021] "Bacteria belonging to the genus Clostridium", which are an active component of the compositions of the invention, are described in the claims.

[0022] Two or more bacterial strains can be used together in the composition of the invention. The use of multiple strains of bacteria belonging to the cluster XIVa or cluster IV in combination may bring an excellent effect on regulatory T lymphocytes. In addition to the bacteria belonging to these clusters, bacteria belonging to other clusters (for example, bacteria belonging to the cluster III) can also be used in combination. When more than one bacterial strain is used (e.g., one or more strains belonging to cluster XIVa, one or more strains belonging to cluster IV, one or more strains belonging to a cluster other than cluster XIVa or cluster IV, such as one or more more strains belonging to cluster III), the type and amount of strains used may vary considerably. The type and amount can be determined depending on various factors (e.g. a desired effect, such as induction or inhibition of regulatory T cell proliferation or accumulation, disease or condition for treatment, prevention or suppression, age and sex of the subject). The strains may be present in one composition, in which case they will be used or consumed together, or they may be present in more than one composition (e.g., each in a separate composition), in which case they may be consumed alone or the compositions may be combined and the resulting the combination (associated compositions) eat or swallow. Any number or combination of strains that will prove effective (e.g., any number from one to 200, e.g., 1 to 100, 1 to 50, 1 to 40, 1 to 30, 1 to 20, 1 to 10, 1 to 5 and a different number between them) can be given. In certain embodiments of the invention, a combination of some or all 46 strains, as described in the document, is used (Itoh, K., and Mitsuoka, T. Characterization of clostridia, and its effect on limited laboratory. Animals 19: 111-118 (1985)). For example, at least one, two or more, three, three or more, four, four or more, five, five or more, six, six or more, or any other number of 46 described strains, including 46 strains, can be used . They can be used in combination with each other or in combination with strains not described in the cited document (e.g., in combination with one or more strains belonging to cluster III). described in the document (Itoh, K., and Mitsuoka, T. Characterization of clostridia). Lab Animals 19: 111-118 (1985)). For example, at least one, two or more, three, three or more, four, four or more, five, five or more, six, six or more, or any other number of 46 described strains, including 46 strains, can be used . They can be used in combination with each other or in combination with strains not described in the cited document (e.g., in combination with one or more strains belonging to cluster III). described in the document (Itoh, K., and Mitsuoka, T. Characterization of clostridia). Lab Animals 19: 111-118 (1985)). For example, at least one, two or more, three, three or more, four, four or more, five, five or more, six, six or more, or any other number of 46 described strains, including 46 strains, can be used . They can be used in combination with each other or in combination with strains not described in the cited document (e.g., in combination with one or more strains belonging to cluster III). 111-118 (1985)). For example, at least one, two or more, three, three or more, four, four or more, five, five or more, six, six or more, or any other number of 46 described strains, including 46 strains, can be used . They can be used in combination with each other or in combination with strains not described in the cited document (e.g., in combination with one or more strains belonging to cluster III). 111-118 (1985)). For example, at least one, two or more, three, three or more, four, four or more, five, five or more, six, six or more, or any other number of 46 described strains, including 46 strains, can be used . They can be used in combination with each other or in combination with strains not described in the cited document (e.g., in combination with one or more strains belonging to cluster III).

[0023] It should be noted that a cluster of "bacteria belonging to the genus Clostridium" may be identified, for example, as follows. Specifically, bacteria belonging to the genus

Clostridia are classified by PCR using a primer set consisting of SEQ ID NOs 64 and 65 (for Clostridium spp. Belonging to cluster XIVa) or a set of primers consisting of SEQ ID NOs 66 and 67 (for Clostridium spp. Belonging to cluster IV) (see Example 18). In addition, bacteria belonging to the genus Clostridium are classified by sequencing the amplified 16S rRNA gene using a primer set consisting of SEQ ID NOS: 19 and 20 (see Example 7).

[0024] Living bacterial cells belonging to the genus Clostridium can be used in the composition of the invention and their killed cells can also be used in the composition. In addition, from the point of view of thermal stability, resistance to antibiotics and the like, and long shelf life, bacteria belonging to the genus Clostridium are beneficial in the form of spore forms.

[0025] The meaning of "physiologically active substance derivative of bacteria belonging to the genus Clostridium" as used herein includes substances contained in bacteria, bacterial secretion products and bacterial metabolites. Such physiologically active substance may be identified by purification of the active ingredient from bacteria, their culture supernatant or the content of the gastrointestinal tract of mice in which only bacteria belonging to the genus Clostridium have been colonized with the already known purification method.

[0026] The active ingredient of the "spore-forming fraction in a mammalian faecal sample" in the composition of the invention is not limited as long as the fraction contains spore-forming bacteria found in the mammalian faeces and has an effect on inducing proliferation or accumulation of regulatory T-lymphocytes.

[0027] The active ingredient "chloroform fraction of a mammalian sample in the composition of the invention is not limited, provided that the fraction is obtained by subjecting the mammal to a mammalian chloroform (e.g., 3% chloroform) and has an effect on induction proliferation or accumulation of regulatory T lymphocytes.

[0028] It should be noted that the "mammal" of the invention is not limited, and examples thereof include humans, mice, rats, horses, cattle, pigs, sheep, monkeys, dogs and cats.

[0029] Meanwhile, when the "spore-forming fraction in a mammalian stool sample" or "chloroform exposed fraction in a mammalian stool sample" is grown in a nutrient medium, bacterial substances, bacterial secretion products, bacterial metabolites are released from bacteria and as such contained in the fraction. The meaning of the active ingredient "fractional culture supernatant" in the composition of the invention includes substances such as secretory products and metabolites. The culture supernatant is not limited, provided that the culture supernatant has an effect on inducing the proliferation or accumulation of regulatory T-cells. Examples of the culture supernatant include the protein fraction of the culture supernatant, the polysaccharide fraction of the culture supernatant,

The composition of the invention may take the form of a pharmaceutical composition, food or drink (which may also be animal feed) or a reagent for experiments.

Over animal models, pharmaceutical compositions, foods or beverages and reagents that affect the induction of regulatory T cell proliferation or accumulation. An example of the invention showed that regulatory T cells (Treg lymphocytes) induced by bacteria and the like belonging to the genus Clostridium weakened proliferation of effector T lymphocytes. Accordingly, the composition of the invention can be advantageously used as a composition having an immunosuppressive effect. The immunosuppressive effect can be assessed, for example, as follows. Specifically, regulatory T cells isolated from an experimental animal, such as a mouse, to which the composition of the invention was orally administered, act on effector T cells (CD4 cells)⁺ CD25^-) isolated from the spleen, after which the proliferation capacity was measured using the dose rate as an indicator [³H] thymidine (see Example 14).

The composition of the invention can be used, for example, in the form of a pharmaceutical composition for the prevention or treatment of autoimmune diseases, such as chronic inflammatory bowel disease, systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis or Hashimoto's disease or allergic diseases, such as like rhinitis or asthma; a pharmaceutical composition for the mitigation of organ transplant rejections and the like; food or drink to improve immune function; or a reagent to attenuate the proliferation or function of effector T cells.

[0032] Further specific examples of target diseases of the compositions of the invention include autoimmune diseases, allergic diseases and organ transplant rejections and the like such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease, arthritis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, transplantation against the host after bone marrow transplantation, osteoarthritis, juvenile idiopathic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, arthritis with spinal joint involvement, systemic lupus erythematosus, insulin-dependent diabetes, thyroiditis, asthma, psoriasis, scleroderma, atopic dermatitis, graft-versus-host disease,acute or chronic immunological diseases related to organ transplantation, sarcoidosis, atherosclerosis, disseminated intravascular coagulation, Kawasaki disease, Graves' disease, nephrotic syndrome, chronic fatigue syndrome, Wegener's granulomatosis, Schonlein-Henoch disease, microscopic renal artery disease, chronic active hepatitis, uveitis, septic shock, toxic shock syndrome, sepsis syndrome, wasting, acquired immunodeficiency syndrome, acute transverse myelitis, Huntington's chorea, Parkinson's disease, Alzheimer's disease, stroke, primary biliary cirrhosis, haemolytic anemia, autoimmune polyglandular syndrome type 1, autoimmune polyglandular syndrome type 2, Schmidt syndrome,syndrome of (acute) respiratory failure (u) of adults, alopecia, alopecia areata, seronegative arthropathy, arthropathy, Reiter's disease, psoriatic arthritis, chlamydia, arthopathy associated with yersinia and salmonella bacteria, ankylosing spondylitis, atherosclerosis, atopic allergy, allergies

-13human, bladder autoimmune disease, pemphigus, pemphigus, pemphigoid, linear IgA bullous dermatosis, autoimmune hemolytic anemia, hemolytic anemia of the positive Coombs reagent, acquired pernicious anemia, juvenile anemia, chronic fatigue syndrome, chronic mucosal candidiasis, giant cell arteritis, primary sclerosing cholangitis, cryptogenic autoimmune hepatitis, acquired immunodeficiency syndrome, acquired immunodeficiency syndrome, hepatitis C, hypogammaglobulinemia, dilated cardiomyopathy, pulmonary fibrosis, cryptogenic fibrosis of the alveolus, post-inflammatory interstitial pneumonitis, interstitial pneumonitis .connective tissue inflammation associated with interstitial pneumonia, mixed connective tissue disease associated with pneumonia, systemic sclerosis associated with interstitial pneumonia, rheumatoid arthritis associated with interstitial pneumonitis, systemic lupus erythematosus associated with pneumonia, dermatomyositis / polymyositis associated with pneumonia, syndromeSjogrenas associated with pneumonia, ankylosing spondylitis associated with pneumonia, pulmonary vasculitis, hemosiderosis associated with pneumonia, drug-induced interstitial lung diseases, radiation-induced lung injury, obliterative bronchiolitis, chronic eosinophilic pneumonia, infiltrative lymphatic inflammation pulmonary, post-inflammatory interstitial pneumonitis, acute gouty arthritis, autoimmune hepatitis, autoimmune hepatitis 1 (classic autoimmune hepatitis), autoimmune hepatitis 2 (hepatitis associated with the presence of anti-LKM antibodies), autoimmune hypoglycemia, insulin-resistant type B with dark keratosis, hypoparathyroidism,acute immune diseases related to organ transplantation, chronic immune disease associated with organ transplantation, osteoarthritis disease, primary sclerosing cholangitis, idiopathic leukopenia, autoimmune neutropenia, NOS kidney inflammation, glomerulonephritis, microscopic systemic renal arteritis, lupus erythematosus, systemic lupus erythematosus, male idiopathic or NOS infertility, autoimmune infertility sperm, multiple sclerosis (all subtypes), insulin dependent diabetes mellitus, sympathetic ophthalmitis, pulmonary hypertension secondary to cellulitis, Goodpasture syndrome, pulmonary signs of periarteritis, acute rheumatic fever , rheumatoid pondos, Still's disease, systemic sclerosis, Takayasu's disease / arteries,autoimmune thrombocytopenia, idiopathic thrombocytopenia, autoimmune thyroid disease, hyperthyroidism, autoimmune hypothyroidism (goitre) (Hashimoto disease), autoimmune hypothyroidism (atrophy), primary myxedema, phakogenetic uveitis, primary vasculitis, vitiligo, allergic rhinitis nose (allergies to pollen), anaphylaxis, allergy to pets, latex allergy, drug allergy, allergic rhinitis, eosinophilic esophagitis, hypereosinophilic syndrome, eosinophilic gastroenteritis, dermal lupusprimary mucopurotic edema, facultative uveitis, primary vasculitis, vitiligo, allergic rhinitis (allergies to pollen), anaphylaxis, pet allergy, latex allergy, drug allergy, allergic rhinitis, eosinophilic esophagitis, syndrome hypereosinophilia, eosinophilic gastroenteritis, cutaneous lupusprimary mucopurotic edema, facultative uveitis, primary vasculitis, vitiligo, allergic rhinitis (allergies to pollen), anaphylaxis, pet allergy, latex allergy, drug allergy, allergic rhinitis, eosinophilic esophagitis, syndrome hypereosinophilia, eosinophilic gastroenteritis, cutaneous lupus

Systemic eosinophilic esophagitis. hypereosinophilic syndrome and eosinophilic gastroenteritis.

The composition according to the invention can be used as a pharmaceutical composition in the prevention or treatment of infectious diseases in an individual whose resistance to infectious diseases is limited due to damage resulting from excessive inflammation caused by immunity.

[0034] Examples of infectious pathogens that reduce the maintenance or recovery of host homeostasis and ultimately cause damage to immunopathological tissues include Salmonella, Shigella, Clostridium difficile, Mycobacterium (which cause tuberculosis), protozoa (which cause malaria), Filarial nematodes (which cause filarioses), Schistosoma (which causes schistosomiasis), Toxoplasma (which causes toxoplasmosis), Leishmania (which causes leishmaniasis), HCV and HBV (which cause hepatitis C and type B) and herpes simplex swiria (which cause herpes simplex).

[0035] Pharmaceutical formulations may be prepared from a composition of the invention by a known method for formulating medicaments. For example, the composition of the invention may be administered orally or parenterally in the form of capsules, tablets, pills, liquids, powders, granules, fine granules, coated preparations, granules, lozenges, sublingual formulations, chewable products, mouthwashes, pastes, suspensions, syrups, elixirs, emulsions, liniments, ointments, patches, wraps, percutaneous absorption systems, fluids, inhalations, aerosols, suppositories, injections and the like.

For the formulation of the preparation, the composition according to the invention can be used in a suitable combination with pharmaceutically acceptable or acceptable carriers for food and beverages, especially sterilized water, physiological saline, vegetable oil, solvent, base material, emulsifier, suspending agent, surfactant, a stabilizer, a fragrance, an aroma, an excipient, a base, a preservative, a binder, a diluent, a hypertonic solution controlling agent, a demulcent, a filler, a disintegrant, a buffer, a coating agent, a lubricant, a coloring agent, a sweetener, a thickening, flavor improving agent, a solubilizing agent, other additives, or the like.

For the formulation of pharmaceutical formulations, especially for the formulation of a pharmaceutical formulation for oral administration, it is advantageous to use in a combination a composition that enables the composition of the invention to be effectively administered to the large intestine, more effectively inducing proliferation or accumulation of regulatory T cells in the large intestine.

[0038] The composition or method of the invention that allows administration to the large intestine is not limited, and known compositions and methods can be used appropriately. Examples include pH sensitive compositions, more specifically enteric polymers,

That release their content when the pH becomes alkaline when the enteric polymers pass through the stomach. When the pH-sensitive composition is used to formulate a pharmaceutical formulation, the pH-sensitive composition should more preferably be a polymer whose threshold pH for the composition is 6.8 to 7.5. Such a range of numerical values ​​is a range in which the pH changes to basic in the distal stomach and is therefore recommended for use in administration to the large intestine.

[0039] In addition, another example of a composition allowing delivery to the large intestine is a composition that provides administration to the large intestine by delaying release of the components for about 3 to 5 hours, which corresponds to a short intestinal transit time. In the formulation of a pharmaceutical formulation using a delayed release composition, a hydrogel is used as a coating. The hydrogel is hydrated and swells after contact with the gastrointestinal juice, thanks to which the content is effectively released. In addition, delayed release dosage units include drug compositions comprising a material that coats or selectively coats the drug. Examples of the selective coating material include degradable in vivo polymers, gradually hydrolysed polymers, polymers that are gradually soluble in water and / or polymers that are degradable in enzymes. A preferred coating material for effective release retardation is not limited, examples include cellulose-based polymers such as hydroxypropylcellulose, acrylic acid polymers and copolymers, such as methacrylic acid polymers and copolymers, and vinylic acid polymers and copolymers, such as polyvinylpyrrolidone.

[0040] Examples of the composition permitting administration to the large intestine include, in addition, bioadhesive compositions that adhere to the colonic mucosa (e.g. a polymer described in US Pat. No. 6,368,686) and compositions into which a protease inhibitor has been included to protect the biopharmaceutical formulation. in the digestive tract before degradation due to protease activity.

[0041] An example of a system allowing delivery to the large intestine is a system for administering the composition of the invention to the large intestine by varying the pressure such that the content is released by using a pressure change caused by gas production in the downstream fermentation process. Such a system is not limited, and its more specific example is a capsule whose contents have been dispersed in the suppository base and which has been covered with a hydrophobic polymer (e.g. ethylcellulose).

[0042] Another example of a system allowing administration to the large intestine is a system for administering compositions to the large intestine, the system being degraded by enzymes (e.g., carbohydrate carbohydrate hydrolases or carbohydrate reductases) present in the large intestine. Such a system is not limited, and its more specific examples include systems that use food components such as non-starch polysaccharides, amylose, xanthan gum and azopolymers.

[0043] When using a pharmaceutical composition, a composition of the invention can be used in combination with already known pharmaceutical compositions for use in immunosuppression. Such a known pharmaceutical composition is not limited and can be at least one therapeutic composition selected from the group consisting of corticosteroids, mesalazine, mesalamine, sulfasalazine, sulfasalazine derivatives, immunosuppressants, cyclosporin A, mercaptopurine, azathiopine, prednisone, methotrexate, antihistamines, glucocorticoids , epinephrine, theophylline, sodium cromoglycate, anti-leukotrienes, anticholinergics for rhinitis, anti-cholinergic decongestants, mast cell stabilizers, anti-IgE monoclonal antibodies, vaccines (preferably vaccines used for vaccination, when the amount of allergen is gradually increased) and combinations thereof. It is preferred to use such therapeutic compositions in combination with the composition of the invention.

[0044] When the composition of the invention is used as a food or beverage, the food or beverage may be, for example, health food, functional food, food for a specific health use, a food supplement, a food for patients or animal feed. The food or beverage can be consumed in the form of a composition according to the invention, and can also be consumed in the form of various food and beverage products. Specific examples of foods and beverages include various beverages, such as juices, refreshing beverages, tea drinks, beverage formulations, gel beverages, and functional beverages; alcoholic beverages, such as beer; carbohydrate-containing foods such as rice-based food products, noodle noodles, bread and pasta; paste products such as hams and fish sausages, seafood pastes; products packed in laminate, plastic or metal foil (retort pouch), such as curry, food with thick starchy sauces, Chinese soups; soups; dairy products, such as milk, milk drinks, ice cream, cheese and yoghurts; fermented products such as fermented soy paste, yoghurts, fermented beverages and marinades; legumes; various confectionery such as western confectionery and biscuits, cakes and the like, Japanese confectionery products, including bean buns with steamed jam, soft adzuki jelly beans and the like, candies, chewing gums, jellies, cold desserts including jelly, creamy caramel and frozen desserts; instant food, such as instant soups and soy-bean soups; microwave reheating foods and the like. Additionally, examples also include healthy foods and drinks prepared in the form of powders, granules, tablets, capsules, liquids, pastes and gels. The composition of the invention can be used in animals, including humans. The use of the composition in non-human animals is not limited, the composition of the invention may be used in a variety of farm animals, poultry, domestic animals, experimental animals and the like. Specific examples of animals include pigs, cattle, horses, sheep, goats, chickens, wild ducks, ostriches, domestic ducks, dogs, cats, rabbits, hamsters, mice, rats, monkeys, and the like, and the list of animals is not limited thereto. including people. The use of the composition in non-human animals is not limited, the composition of the invention may be used in a variety of farm animals, poultry, domestic animals, experimental animals and the like. Specific examples of animals include pigs, cattle, horses, sheep, goats, chickens, wild ducks, ostriches, domestic ducks, dogs, cats, rabbits, hamsters, mice, rats, monkeys, and the like, and the list of animals is not limited thereto. including people. The use of the composition in non-human animals is not limited, the composition of the invention may be used in a variety of farm animals, poultry, domestic animals, experimental animals and the like. Specific examples of animals include pigs, cattle, horses, sheep, goats, chickens, wild ducks, ostriches, domestic ducks, dogs, cats, rabbits, hamsters, mice, rats, monkeys, and the like, and the list of animals is not limited thereto.

[0045] Without referring to the theory, individuals in whom the relative abundance of bacteria belonging to the Firmicutes group (the group to which Clostridium clusters IV and XIVa belong) are large, gain greater mass gain, than individuals in whom the relative abundance of bacteria belonging to the Bacteroidetes group is large. Accordingly, the composition according to the invention may condition the absorption of nutrients and the improvement of feed efficiency. From this point of view, the composition of the invention can be used to improve weight gain or as animal feed with good yield.

[0046] In addition, the addition of a composition according to the invention to the non-antibiotic feed makes it possible to increase the weight of the individual who eats the feed at a level equal to or higher than the mass achieved with the intake of the feed containing the antibiotics and also makes it possible to reduce the pathogenic bacteria in the stomach, at a level equal to that obtained with the consumption of a typical feed containing antibiotics. Accordingly, the composition of the invention can be used in animal feeds that do not require the addition of antibiotics.

[0047] In addition, for commercial purposes, in contrast to traditional bacteria (Lactobacillus and Bifidobacteria), which are not easy to use in animal production, the composition of the invention in the form of spore forms can be used in tablets, sprayed or mixed with feed, and can also be added to drinking water.

Feeding the composition according to the invention is not limited and the feed may be administered to the subject at regular intervals, selectively or may be administered for some time (e.g., at birth, during weaning or when the fed individual is to be transferred or sent elsewhere).

[0049] In addition, the composition of the invention can be used in malnourished people. In other words, also when the individual who adopts the composition is a human, the composition can be used to promote weight gain and increase energy absorption from food.

[0050] The food or drink can be produced using a manufacturing technique that is well known in the art. One or more components (e.g., nutrients) can be added to the food or beverage, which effectively improve the immune system's function by immunosuppressive action. In addition, the food or drink may be combined with another ingredient or other functional food having functions other than the function of improving the immune function, thus serving as a multi-functional food or beverage.

[0051] In addition, the composition of the invention may be added to food products that require a treatment step that can destroy common probiotic strains. Specifically, most commercial useful probiotic strains can not be added to foods that are to be subjected to any heat treatment, long-term storage, freezing, processing involving mechanical stresses and high pressure treatment (e.g.

- embossing or rolling). On the other hand, due to the advantageous nature of the spore forms, the composition according to the invention can be added without problem to processed foods.

[0052] For example, the compositions of the invention in the form of spore forms can survive even in dried foods and can remain viable even after ingestion. Similarly, the composition of the invention can withstand low temperature sterilization processes, typically processes at a temperature in the range of 70 ° C to boiling point, both inclusive. The composition according to the invention can be added to all types of dairy products. In addition, the composition of the invention can withstand long-term, long-term storage; high temperature treatment, such as cooking and baking; low temperature processing, such as freezing and cooling; and high pressure treatment such as extrusion molding and rolling.

[0053] Food products that need to be processed under such difficult conditions are not limited and examples thereof include food products that have to be treated in a microwave oven to be edible (e.g., porridge), food that needs to be washed to ensure it was edible (for example, muffins), food which should be subjected to sterilization at high temperature in a short time to be edible (e.g., milk) and foodstuffs that need to be heated to be drinkable (e.g., hot tea ).

When the composition according to the invention is administered or consumed, its amount to be administered or ingested is appropriately selected depending on the age, body weight, symptoms, health status of the subject, type of composition (pharmaceutical product, food or drink or the like) and this similar. For example, the amount to be administered or ingested is usually 0.01 mg / kg body weight to 100 mg / kg body weight, preferably 1 mg / kg body weight to 10 mg / kg body weight.

[0055] Also described is a product of the composition of the invention (pharmaceutical product, food or drink or reagent) or description thereof that can be provided with information that the product can be used to attenuate immunity (including information that the product has immunosuppressive activity and information that the product has the effect of inhibiting the proliferation or function of effector T lymphocytes). Here, the "delivery of a product or its description with information" means that information has been included on the main body, container, packaging or similar product, or the information is included in the description, leaflet or other printed material that discloses the product information.

<Method of induction of proliferation or accumulation of regulatory T lymphocytes>

[0056] As described above and as set forth in the Examples, administration of a composition of the invention to a subject allows induction of regulatory T cell proliferation or accumulation in a subject.

[0057] It should be noted that the term "subject" used is not limited, examples thereof include humans, various types of cattle, poultry, pets, animals

-19 experimental and the like. An "individual" may be in a state of health or in a state of illness.

[0058] In addition, as will be shown in Example 5, gram-positive commensal bacteria play a major role in the proliferation or accumulation of regulatory T-cells.

The term "antibiotic against gram-negative bacteria" as used herein is not limited, and examples include aminoglycoside antibiotics (amikacin, gentamycin, kanamycin, neomycin, netylmycin, tobramycin and paromomycin), cephalosporin antibiotics (cefaclor, cefamandol, cefoxitin, cefprozil, cefuroxime , cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone and cefoxitin), sulfonamides, ampicillin and streptomycin. Without referring to the theory, the "antibiotic against Gram-negative bacteria" according to the invention is preferably one that reduces the number of gram-negative bacteria and also contributes to the colonization of gram-positive bacteria.

In addition, the components of the probiotic, such as almond peel, inulin, oligofructose, raffinose, lactulose, pectin, hemicellulose (e.g. xyloglucan and alpha-glucans), amylopectin and resistant starch, do not decompose in the upper gastrointestinal tract and affect on the growth of intestinal microbes in the digestive tract, as well as growth factors, such as acetyl-CoA, biotin, beet molasses and yeast extracts, contribute to the proliferation of bacteria belonging to the genus Clostridium.

[0061] The "anti-gram-negative antibiotic" and "probiotic composition or growth factor" described above can be used in combination. Such combined use is not limited, examples of combined use are the following: "an antibiotic against Gram-negative bacteria" is administered to a subject earlier, after which a composition of the invention is administered; An "antibiotic against gram-negative bacteria" and the composition of the invention is administered to the subject simultaneously; A "probiotic composition or growth factor" is administered to a subject earlier, after which a composition of the invention is administered; "Probiotic composition or growth factor" and the composition of the invention is administered to the subject simultaneously; a composition according to the invention,

[0062] In addition, the therapeutic composition may be administered to the subject together with at least one substance selected from the group consisting of the composition of the invention, "antibiotic against Gram-negative bacteria" and "probiotic or growth factor composition".

[0063] Such a therapeutic composition is not limited and may be at least one therapeutic composition selected from the group consisting of corticosteroids, mesalazine, mesalamine, sulfasalazine, sulfasalazine derivatives, immunosuppressive drugs, cyclosporin A, mercaptopurine, azathiopine, prednisone, methotrexate, antihistamines , glucocorticosteroids, epinephrine, theophylline, sodium cromoglycate,

Anti-leukotrienes, anticholinergics for rhinitis, anticholinergic antiglaucoma drugs, mast cell stabilizers, anti-IgE monoclonal antibodies, vaccines (preferably vaccines used for vaccination in the case where the amount of allergen is gradually increased) and combinations thereof. The use of therapeutic compositions in combination with the described substances is preferred.

[0064] In addition, there is no restriction on the combined use of the therapeutic composition with at least one substance selected from the group consisting of the composition of the invention, "antibiotic against Gram-negative bacteria" and "probiotic or growth factor composition". For example, "one substance" and a therapeutic composition are administered to the subject orally or parenterally simultaneously or separately at any appropriate time.

[0065] In addition, in a "method of inducing proliferation or accumulation of regulatory T lymphocytes," it can be determined whether the use of a composition of the invention or the like actually induces the proliferation or accumulation of regulatory T lymphocytes using, as an indicator, an increase or reinforcement of at least one member from a selected compound group from the number of regulatory T lymphocytes, the ratio of quantitative regulatory T cells in the group of colon T lymphocytes, the function of regulatory T lymphocytes and the expression of a regulatory T-cell marker. It is preferred to use one measurement selected from the group consisting of stimulation of IL-10 expression, stimulation of CTLA4 expression, stimulation of IDO expression and inhibition of IL-4 expression as an indicator of induction of proliferation or accumulation of regulatory T-lymphocytes.

[0066] It should be noted that examples of the expression detection method include northern blot, real-time RT-PCR and dot blot hybridization for detecting gene expression at the transcription level; and an ELISA, radioimmunoassays, immunoassay, immunoprecipitation and flow cytometry to detect gene expression at the translation level.

[0067] The sample used to measure the index is not limited, examples thereof include taking blood from the subject and fragments of tissues obtained as a result of biopsy.

<A method of predicting the response of an individual to a composition of the invention and / or prognosis of an individual>

[0068] A method is described wherein the total amount or the ratio of bacteria belonging to the genus Clostridium to the subject's microbiota is determined, and when the ratio or absolute value of the bacteria belonging to the genus Clostridium is reduced relative to the original value obtained in determining the individual in the subject a typical health condition has been established that the subject probably responds to the composition of the invention.

[0069] A method for predicting a subject's response to substance and / or prognosis of a subject is provided herein. The method comprises measuring the percentage or absolute value of clostridium IV and XIV clusters in the subject's microbiota and comparing with the starting value of the same measurements in the prototype healthy subject, where the depression

The absolute or percentage level of clusters IV and / or XIV of Clostridium indicates that the subject may preferably respond to the composition of the invention.

[0070] The method may further include measuring the composition of the microbiota of the subject after administration of the substance, wherein an increase of the percentage or absolute value of the species Clostridium belonging to the cluster IV, XIV, when the composition of the invention in relation to the measurements before administration is a positive indicator of increased immunosuppression (or immunoregulation). Measurement of the composition of the subject's microbiota may be performed using techniques known in the art, such as 16SrRNA sequencing.

[0071] It should be noted that in these methods the substance is at least one substance selected from the group consisting of the following substances (a) to (e):

(a) bacteria belonging to the genus Clostridium or a physiologically active substance derived from bacteria;

(b) a fraction forming spores forms of a stool sample obtained from a mammal or a culture supernatant of a fraction;

(c) a chloroform-treated fraction of a stool sample obtained from a mammal or a culture supernatant of a fraction;

(d) antibiotic against Gram-negative bacteria; and (e) at least one substance selected from the group consisting of almond peel, inulin, oligofructose, raffinose, lactulose, pectin, hemicellulose (e.g., xyloglucan and alpha-glucans), amylopectin, acetyl CoA, biotin, beet molasses, extract yeast and resistant starch.

<Method of inhibiting the proliferation or accumulation of regulatory T lymphocytes>

[0072] As shown in Example 5, gram-positive commensal bacteria play a major role in the proliferation or accumulation of regulatory T lymphocytes. Accordingly, a method is described for inhibiting the proliferation or accumulation of regulatory T cells in a subject, the method comprising the step of administering to the subject an antibiotic against gram-positive bacteria.

[0073] The term "antibiotic against gram-positive bacteria" as used herein is not limited, examples thereof include cephalosporin antibiotics (cefalexin, cefuroxime, cefadroxil, cefazolin, cefalotin, cefaclor, cefamandol, cefoxitin, cefprozil and ceftobiprol); fluoroquinolone antibiotics (ciprofloxacin, Levaquin, ofloxacin, gatifloxacin (tequin), moxifloxacin and norfloxacin); tetracycline antibiotics (tetracycline, minocycline, oxytetracycline and doxycycline); penicillins (amoxicillin, ampicillin, penicillin V, dicloxacillin, carbenicillin, vancomycin and methicillin) and carbapenem antibiotics (ertapenem, doripenem, imipenem / clastatin and meropenem).

[0074] The term "subject" used in the invention is not limited, examples thereof include humans, various types of cattle, poultry, domestic animals, experimental animals and the like. An "individual" may be in a state of health or in a state of illness. The condition of the disease

It is not limited, examples thereof include those subject to cancer immunotherapy and an infectious disease.

[0075] In addition, another "a method to inhibit proliferation or accumulation of regulatory T cell" is a method for inhibiting proliferation or accumulation of regulatory T cells in a subject, comprising the step of administering to the subject one of the antibodies, fragment of an antibody and a peptide which act against the antigen which is at least one substance selected from the group consisting of the following substances (a) to (c):

(a) bacteria belonging to the genus Clostridium or a physiologically active substance derived from bacteria;

(b) a fraction forming spores forms of a stool sample obtained from a mammal or a culture supernatant of a fraction;

(c) a chloroform-treated fraction of a stool sample obtained from a mammal or a culture supernatant of a fraction;

<Vaccine composition and method for the treatment or prevention of infectious diseases or autoimmune diseases using a vaccine composition>

[0076] As shown in Example 15, the induction of lymphocyte Tregs in the large intestine by Clostridium plays an important role in local and systemic immune responses. Accordingly, also described is a "vaccine composition comprising at least one substance selected from the group consisting of the following substances (a) to (c): (a) bacteria belonging to the genus Clostridium; (b) spore forms of bacteria in the fraction forming the spore forms of the stool sample obtained from the mammal; (c) bacteria of the chloroform fraction treated with a chloroform stool sample obtained from a mammal "and" a method of treating, aiding a treatment, diminishing or preventing at least one disease selected from infectious diseases and autoimmune diseases in a subject, a method comprising administering a vaccine composition to a subject. "

[0077] It should be noted that "autoimmune diseases" are not limited, examples thereof include those described in "specific examples of target diseases" in a <RTI ID = 0.0> composition </ RTI> inducing proliferation or accumulation of regulatory T lymphocytes>. "Infectious diseases" are also not limited, examples thereof include infectious diseases associated with "infectious pathogens" described as "examples of infectious pathogens" in a <inducing composition of proliferation or accumulation of regulatory T lymphocytes>.

<A method of screening a compound with an effect contributing to the proliferation or accumulation of regulatory T lymphocytes>

[0078] Also described is screening of a compound having an action contributing to the proliferation or accumulation of regulatory T-cells, which include:

(1) the preparation of a test substance from at least one substance selected from the group consisting of the following substances (a) to (c):

(A) bacteria belonging to the genus Clostridium or a physiologically active substance of a bacterium derivative;

(b) a fraction forming the spore forms of a stool sample obtained from a mammal or culture supernatant fraction; and (c) a chloroform-treated fraction of a stool sample obtained from a mammal or a supernatant of a cultured fraction.

(2) the preparation of non-human mammals in which the reporter gene is expressed under the control of IL-10 gene expression;

(3) administration of the test substance to non-human mammals;

(4) after administration of the test substance, detection of cells expressing the reporter gene in the CD4 cell group⁺ Foxp3⁺ a non-human mammal, and determining the number of cells in the CD4 cell group⁺ Foxp3⁺ expressing the reporter gene or the cell volume ratio in the CD4 cell group⁺ Foxp3⁺ expressing the reporter gene into cells in the CD4 cell group⁺ Foxp3⁺ not expressing the reporter gene;

(5) detection of cells expressing the reporter gene in the CD4 cell group⁺ Foxp3⁺ in non-human mammals that were not given the test substance and determining the number of cells in the CD4 cell group⁺ Foxp3⁺ expressing the reporter gene or the quantitative ratio of cells from the CD4 cell group⁺ Foxp3⁺wykazujących ekspresję genu reporterowego do komórek w grupie komórek CD4⁺ Foxp3⁺ not expressing the reporter gene; and (6) a comparison of the magnitude or ratios specified in (4) with the value or quantitative ratio specified in (5) and determining when the value or ratio specified in (4) is greater than that specified in (5), so that the test substance is a compound beneficial for the proliferation or accumulation of Treg lymphocytes.

[0079] The term "test substance" is not limited, provided that the test substance is a substance made of at least one substance selected from the group consisting of a substance (a) to (c). Examples of a test substance include proteins, polysaccharides, fats and nucleic acids that are derived from at least one substance selected from the group consisting of the above-described substances (a) to (c).

[0080] The term "non-human mammal in which the reporter gene is expressed during the control of IL-10 gene expression" is not limited, provided that the non-human mammal is a non-human mammal having a reporter gene whose expression is region-controlled. control of IL-10 gene expression (e.g., a gene promoter or enhancer). Examples of such reporter genes include genes encoding fluorescent proteins (e.g., GFP) and luciferase-encoding genes. As a "non-human mammal in which the reporter gene is expressed under the control of IL-10 gene expression" according to the invention, an Il10 mouse may be preferably used.^venusas shown in the examples.

[0081] The term "administration / administration" used in the invention is not limited, examples thereof include administering the test substance to non-human orally or parenterally administrable animals (e.g., intraperitoneal injections or intravenous injections).

[0082] A non-human mammal has been described that has been used in a method whose reporter gene is expressed under the control of IL-10 gene expression.

[0083] In addition, a method is also described for isolating bacteria belonging to the genus Clostridium from a sample, a compound having an activity promoting the proliferation or accumulation of regulatory T lymphocytes, the method comprising the following steps (1) to (3):

(1) preparation of genomic DNA from a sample of bacteria belonging to the genus Clostridium;

(2) placing genomic DNA in a cloning system and producing a library of genes derived from a sample of bacteria belonging to the genus Clostridium; and (3) isolating a compound having an activity that promotes the proliferation or accumulation of regulatory T lymphocytes by using the gene library obtained in step (2).

[0084] In these steps, the preparation and isolation methods are not limited and known in vitro or in vivo techniques can be used as appropriate. In addition, the compound isolated by this method is not limited, examples thereof include nucleic acids (e.g. DNA, mRNA and rRNA) derived from bacteria belonging to the genus Clostridium as well as polypeptides and proteins derived from bacteria belonging to the genus Clostridium.

[0085] Also described is a method for determining the composition of a microbiota in an individual where an increase in the ratio or total number of bacteria belonging to the genus Clostridium after administration of a composition of the invention to a subject having a quantitative or absolute ratio prior to administration has been used as an indicator of increased immunosuppression. In such a method, the method for determining the composition of a microbiota is not limited and known techniques (e.g., 16S rRNA sequencing) can be used as appropriate.

[0086] Also described is a method for measuring Treg differentiation in lymphocytes, where an increase in Treg differentiation in a subject after administration of a composition of the invention including pre-dose values ​​has been used as an indicator of increased immunosuppression (or immunoregulation).

[0087] In addition, the composition of the invention may be administered to the subject during antibiotic treatment. The time intervals of administration are not limited and the composition can, for example, be administered before or simultaneously with antibiotics. The composition should preferably be administered in the form of spore forms from the point of view of resistance to antibiotics.

[0088] In addition, in a preferred mode of administration, the composition of the invention is administered after or simultaneously with, for example, an antibiotic against Gram-positive bacteria. It should be noted that the "antibiotic against gram-positive bacteria" is not limited, examples thereof include cephalosporin antibiotics (cefuroxime, cefalexin, cefadroxil, cefazolin, cefalotin, cefaclor, cefamandol, cefoxitin, cefprozil and ceftobiprol); fluoroquinolone antibiotics (ciprofloxacin, Levaquin, ofloxacin, gatifloxacin (tequin), moxifloxacin and norfloxacin); tetracycline antibiotics (tetracycline, minocycline, oxytetracycline and doxycycline); penicillins (amoxicillin, ampicillin, penicillin V, dicloxacillin, carbenicillin, vancomycin and methicillin) and carbapenem antibiotics (ertapenem, doripenem, imipenem / clastatin and meropenem).

[0089] Simultaneously, in another preferred method of administration, the composition of the invention is administered after (or simultaneously) treatment using, for example, vancomycin, metronidazole, linezolid, ramoplanin or fidaxomycin.

[Examples] [0090] Hereinafter, the invention will be described in more detail by way of examples. However, the invention is not limited to the following examples.

[0091] It should be noted that the mice used in the examples were prepared or created in the following way. In the description, mice may be referred to as "SPF" or "GF". "SPF" and "GF" mean that mice were properly housed in conditions free from specific pathogenic bacteria (free from specific pathogens, SPF) and that mice were kept in a germ-free (GF) condition.

<Mouse>

[0092] C57BL / 6, Balb / ci IQI mice housed in SPF or GF conditions were purchased from Sankyo Labo Service Corporation, Inc. (Japan), Japan SLC, Inc. (Japan), CLEA Japan, Inc. (Japan) or The Jackson Laboratory (USA). GF mice and gnotobiotic mice were reproduced and kept in the gnoxotic facilities of the University of Tokyo, the Yakult Central Institute for Microbiological Research or Sankyo Labo Service Corporation, Inc. Myd88 mice^{- / -}, Rip2^{- / -}, and Card9^{- / -} grown, as described in non-patent documents up to 3, backcrossed for 8 generations or more, and genetic background achieved

C57BL / 6. Foxp3 mice^eGFP was purchased from the Jackson Laboratory.

<Il10 Mice^venus>

[0093] To create a bicistronic locus encoding both Il10 and Venus under the control of the Il10 promoter, the target construct was created. Specifically, a cassette (cassette with IRESVenus-SV40-polyA signal, see Non-Patent Document 4), which was created from internal ribosomal binding (IRES), yellow fluorescent protein (Venus) and SV40 PoliA signal (SV40 poliA) and placed next to the gene (neo) neomycin resistant, inserted between the stop codon and the polyA signal (exon 5) of the Il10 gene. Then, the resulting target construct was used for homologous recombination with the Il10 gene region in the mouse genome. Thus, Il10 mice were created^venushaving an Il10 allele^venus (see Fig. 1).

It should be noted that in Figure 1 "tk" is a gene encoding thymidine kinase, "neo" means

Neomycin-resistant resistance and "BamH1" means the cleavage site of the BamH1 restriction enzyme.

[0094] Genomic DNA was extracted from Il10 mice^venus, treated with BamHI and subjected to Southern blotting using the probe shown in Fig. 1. Fig. 2 shows the results obtained. Wild type alleles and Il10 alleles were detected^venus as bands of 19 kb and 5.5 kb respectively. As shown by the results shown in Fig. 2, it was found that the homologous recombination shown in Fig. 1 occurred in the genome of Il10 mice.^venus.

[0095] In addition, CD4 cells⁺ venus^- or CD4 cells⁺ venus⁺ in the lamina propria of the large intestine of Il10 mice^venus sorted using FACSAria. Real-time RT-PCR was then performed on the ABI 7300 system, as described below, to determine the amount of IL-10 mRNA expressed. Figs. 3 and 4 show the results obtained. As can be seen from the results presented in Figs. 3 and 4, it was found that the development of IL-10 mRNA was only detected in CD4 cells⁺ venus⁺, expression of IL-10 mRNA in Il10 mice^venus was correctly reflected in the Venus expression. It should be noted that Il10 mice^venus free of microbes was created at the Central Institute for Experimental Animals (Kawasaki, Japan). Il10 mice^venus microbial-free holders were kept in vinyl insulators at Sankyo Labo Service Corporation, Inc., (Tokyo, Japan) and used in the following examples.

[0096] The experiments and analyzes in the examples were carried out as follows.

<Method of colonization of mice with bacteria and its analysis>

[0097] As described in Non-patent Documents 5 and 6, mice that were colonized with SFB or Clostridium were cultured. The contents of the cecum or faeces obtained from gnotobiotic mice were dissolved in sterile water or in an anaerobic dilution solution. The dissolved contents of the cecum or dissolved faeces, optionally after treatment with chloroform, were orally administered to GF mice. Three strains of Lactobacillus and 16 Bacteroides strains were grown anaerobically, each separately, in BL or EG agar medium. The cultured bacteria were harvested, suspended in anaerobic TS medium and administered by gavage to GF mice. The colonization status of bacteria in mice was assessed by microscopic observation of faecal smear.

<Cell separation and flow cytometry>

[0098] For the isolation of lymphocytes from the lamina propria of the large intestine and the lamina propria of the small intestine, the small intestine and large intestine were obtained and dissected longitudinally. Next, the content of stool and the like was removed from the inside by rinsing. The small intestine and large intestine were then shaken in HBSS containing 5 mM EDTA at 37 ° C for 20 minutes. After removal of the epithelium and adipose tissue, the intestinal tissues were cut into small pieces. RPMI 1640 (4% fetal bovine serum (FBS), 1 mg / ml collagenase D, 0.5 mg / ml dispase and 40 μg / ml DNaseI (all manufactured by Roche Diagnostics KK) were added to small pieces), and the mixture was shaken in bath at 37 ° C for 1 hour. The digested tissues were washed in HBSS containing 5 mM EDTA and again

Was suspended in 5 ml of 40% Percoll (GE Healthcare). A layer of 2.5 ml 80% Percoll in a 15-ml Falcon tube was applied to the suspension. It was then centrifuged at room temperature at 2000 rpm for 20 minutes to separate the cells by Percoll density gradient centrifugation. Cells were collected at the interface and used as lamina propria. Harvested cells were suspended in staining buffer (PBS, 2% FBS, 2 mM EDTA and 0.09% NaN3) and stained using anti-CD4 antibody (RM4-5, BD Biosciences), labeled using PE or PE-Cy7. Upon completion of CD4 staining, Foxp3 was stained in cells using Cytofix / Cytoperm Kit Plus from Golgistop (BD Biosciences) or Foxp3 Staining Buffer Set (eBioscience) as well as Foxp3 antibodies (FJK-16s, eBioscience) labeled using Alexa647. Flow cytometry was performed using FACScant II, and the data was analyzed using FlowJo software (TreeStar Inc.). Cell sorting was performed using FACSAria.

<RT-PCR in real time>

[0099] From RNA prepared using the RNeasy Mini Kit (Qiagen), the cDNA was synthesized using MMV reverse transcriptase (Promega KK). The resulting cDNA was analyzed in RT-PCR in real time using Power SYBR Green PCR Master Mix (Applied Biosystems) and ABI 7300 real-time RT-PCR system (Applied Biosystems) or RT-PCR in real time using SYBR Premix Ex Taq (TAKARA ) and Light Cycler 480. For each sample, the obtained value was normalized to the amount of GAPDH. The primer set was prepared using Primer Express version 3.0 (Applied Biosystems) and those showing 90% or

<td>greater sequence identity during the initial evaluation. the starter was as follows:</td><td>Set</td><td colspan="2">applied</td>

<td>Foxp3 5'-GGCAATAGTTCCTTCCCAGAGTT-3 '(SEQ 5'-GGGTCGCATATTGTGGTACTTG-3 '(SEQ ID NO: 2)</td><td>ID</td><td>NO:</td><td>1)</td>

<td>CTLA4 5'-CCTTTTGTAGCCCTGCTCACTCT-3 '(SEQ 5'-GGGTCACCTGTATGGCTTCAG-3 '(SEQ ID NO: 4)</td><td>ID</td><td>NO:</td><td>3)</td>

<td>GITR 5'-TCAGTGCAAGATCTGCAAGCA-3 '(SEQ 5'-ACACCGGAAGCCAAACACA-3 '(SEQ ID NO: 6)</td><td>ID</td><td>NO:</td><td>5)</td>

<td colspan="2">IL-10 5'-GATTTTAATAAGCTCCAAGACCAAGGT-3 '(SEQ ID 5'-CTTCTATGCAGTTGATGAAGATGTCAA-3 '(SEQ ID NO: 8)</td><td>NO:</td><td>7)</td>

<td>GAPDH 5'-CCTCGTCCCGTAGACAAAATG-3 '(SEQ 5'-TCTCCACTTTGCCACTGCAA-3 '(SEQ ID NO: 10)</td><td>ID</td><td>NO:</td><td>9)</td>

<td>Mmp2 5'-GGACATTGTCTTTGATGGCA-3 '(SEQ 5'-CTTGTCACGTGGTGTCACTG-3 '(SEQ ID NO: 12)</td><td>ID</td><td>NO:</td><td>11)</td>

<td>Mmp9 5'-TCTCTGGACGTCAAATGTGG-3 '(SEQ 5'-GCTGAACAGCAGAGCCTTC-3 '(SEQ ID NO: 14)</td><td>ID</td><td>NO:</td><td>13)</td>

<td>MMP13</td><td>AGGTCTGGATCACTCCAAGG 5'-3 '</td><td>(KNOT</td><td>ID</td><td>NO:</td><td>15)</td>

<td colspan="2">5'-TCGCCTGGACCATAAAGAA-3 '(SEQ ID NO: 16)</td><td></td><td></td><td></td><td></td>

<td>Ido1</td><td>AGAGGATGCGTGACTTTGTG 5'-3 '</td><td>(KNOT</td><td>ID</td><td>NO:</td><td>17)</td>

5'-ATACAGCAGACCTTCTGGCA-3 '(SEQ ID NO: 18).

<Preparation and culture of colon epithelial cells (IEC)>

[0100] First, the colon was obtained, dissected longitudinally and rinsed with PBS. The colon was then treated with 1 mM dithiothreitol (DTT) at 37 ° C for 30 minutes on a shaker and then centrifuged for one minute to break the integrity of the epithelium. The released IECs were collected and resuspended in 5 ml of 20% Percoll. A layer of 2.5 ml 80% Percoll was applied to the suspension in a 15 ml Falcon tube. The tubes were then centrifuged at 25 ° C and 780 g for 20 minutes to separate the cells by Percoll density gradient centrifugation. Cells were harvested at the interface and IEC large intestine was used (purity: 90% or higher, viability: 95%). Obtained IEC was suspended in RPMI containing 10% FBS and 1 x 10⁵ IEC cells were grown in a 24-well plate for 24 hours. The culture supernatant was then collected and the level of active TGFβ1 measured by ELISA (Promega).

[0101] At the same time, 1.5 x 10 were grown in vitro for the growth of T lymphocytes⁵ MACS of purified CD4 T-lymphocytes⁺ spleen in each round bottom well of a 96-well plate along with 50% conditioned medium containing IEC isolated from GF mice or colonized mice with Clostridium together with 25 ng / ml hIL-2 (Peprotech), in the presence or absence of 25 μg / ml of anti-TGF-antibodies β (R &amp; D). It should be noted that 10 μg / ml of anti-CD3 antibodies and anti-CD28 antibodies (BD Bioscience) were plated on a round bottom plate. After 5 days culture, CD4 T-lymphocytes⁺collected and tested using real-time PCR.

<Experimental model of colitis>

[0102] A faecal suspension of mice colonized with Clostridium was orally administered to C57BL / 6 mice (2-week old) and cultured in a conventional environment for six weeks.

For the preparation of the induced DSS model of colitis, 2% (w / v) DSS (pure reagent, DSS salt, molecular weight = 36 to 50 kD, produced by MP Biomedicals) was administered to mice with drinking water for six days .

[0104] At the same time, for the preparation of oxazolone-induced colitis model, mice were previously sensitized to oxazolone percutaneously, 150 μl 3% oxazolone (4-ethoxymethylene-2-phenyl-2-oxazolin-5-one, Sigma-Aldrich) / 100% ethanol solution. Five days later, 150 μl of 1% oxazolone / 50% ethanol solution was rectally administered once more with previously sensitized mice under mild anesthesia. It should be noted that rectal administration was performed using a 3.5F catheter.

[0105] Each mouse was examined daily for body weight, latent blood, bleeding visible to the naked eye (gross blood) and stool hardness. In addition, the percentage ratio

Loss of weight, intestinal bleeding (no bleeding, blood in the stool (hemoccult +) or bleeding visible to the naked eye) and stool hardness (normal stool, loose stool or diarrhea) were quantified and the disease activity index (DAI) calculated as described in " S. Wirtz, C. Neufert, B. Weigmann, MF Neurath, Nat Protoc 2, 541 (2007). "

<Specific IgE response to OVA>

[0106] BALB / c SPF mice were inoculated with suspensions of faeces of Clostridium (2-week) colonized mice and cultured in a conventional environment. Then, 1 μg of OVA (class V, Sigma) at a dose of 2 mg alum (Thermo Scientific), 0.2 ml total, was injected intraperitoneally into mice (when they reached the age of 4 weeks and 6 weeks). Weekly, sera were harvested from mice at the base of the tail and specific OVA IgE was measured by ELISA (Chondrex). Then, when the mice reached the age of 8 weeks, the spleen cells were harvested and inoculated in a 96-well plate at a concentration of 1 x 10⁶ cells per well and stimulated with OVA (100 μg / ml) for three days. The culture supernatant was then collected and the levels of IL-4 and IL-10 were measured by ELISA (R &amp; D).

<Statistical analysis>

[0107] Differences between control and experimental groups were evaluated using Student's t-test.

(Example 1) [0108] First, it was examined whether the accumulation of regulatory T lymphocytes (Treg lymphocytes) in the lamina propria of the intestine was dependent on commensal bacteria. Specifically, lymphocytes from peripheral lymph nodes (pLN) of Balb / c mice were isolated in the absence of specific pathogenic bacteria (SPF) or from the lamina propria or small intestine (SI) of mice. CD4 and Foxp3 were stained with antibodies. The ratio of Foxp3 was then analyzed⁺ in CD4 cells⁺ flow cytometry. Fig. 5 shows the obtained results. As can be seen from the results presented in Fig. 5, it was found that Trep Foxp3 lymphocytes⁺ they are present at high frequency in the lamina propria of the gastrointestinal tract, especially in the lamina propria of the large intestine of mice kept in an environment free of specific pathogenic microorganisms (SPF). In addition, it was also found that the number of Treg Foxp3 lymphocytes⁺ in the lamina propria of the large intestine gradually increases up to three months after birth, and the number of lymphocytes Treg Foxp3⁺ in peripheral lymph nodes is essentially constant from the moment after the second week after birth.

(Example 2) [0109] Next, it was investigated whether the temporal accumulation of Treg cells in the large intestine, as determined in Example 1, is associated with colonization of the intestines with commensal microbiota. Specifically, the expression of CD4 and expression of Foxp3 in lymphocytes isolated from the small intestine, large intestine and peripheral lymph nodes of mice cultured in a microbial-free (GF) or SPF environment (age 8 weeks: Balb / c mice, IQI mice and C57BL mice) were analyzed. 6). Similar results were obtained in three or more

-Non-independent studies. Fig. 6 and 7 show the obtained results. It should be noted that in Figure 7 each white circle represents the absolute value of CD4 lymphocytes⁺ Foxp3⁺ in each mouse, and error bars are standard deviations (SD).

[0110] In addition, lymphocytes from the SPF mice and GF mice (BALB / c mice or C57BL / 6 mice) were collected. CD4 and Foxp3 were stained with antibodies. The lamina propria lymphocytes were then analyzed using FACS. Fig. 8 shows the results obtained. It should be noted that in Figure 8 each white circle represents the absolute value of CD4 cells⁺ Foxp3⁺ in each mouse, ** means "P &lt;0.001" and a * means "P &lt;0.01".

[0111] In addition, lymphocytes were isolated from the lamina propria of the large intestine, the lamina propria of the small intestine (SI), Peyer's patches (PP) and mesenteric lymph nodes (MLN) of SPF mice (C57BL / 6 mice), which were orally administered antibiotics with water for eight weeks . CD4 and Foxp3 were stained with antibodies. Lymphocytes were then analyzed using FACS. Similar results were obtained in two or more independent studies. Fig. 9 shows the results obtained (the ratio of Foxp3 cells⁺ in CD4 cells⁺ in each mouse separately). It should be noted that the following antibiotics were used in combination as described in the document:

ampicillin (A, 500 mg / l, Sigma) vancomycin (V, 500 mg / l, NACALAI TESQUE, Inc.) metronidazole (M; 1 g / l, NACALAI TESQUE, Inc.) neomycin (N; 1 g / l, NACALAI TESQUE, Inc.) [0112] Rakoff-Nahoum, J. Paglino, F. Eslami-Varzaneh, S. Edberg, R. Medzhitov, Cell 118, 229 (Lip 23, 2004) [0113] Fagarasan et al., Science 298, 1424 (Nov. 15, 2002) [0114] In Fig. 9, each white circle represents the absolute value of CD4 cells⁺ Foxp³+ in each mouse separately, each horizontal bar represents the mean value of the absolute values, * means that "P &lt;0.01" and "AVMN" means the types of antibiotics administered identified by the first antibiotic letter.

[0115] As can be seen from the results shown in Figures 6 to 9, the frequency and absolute values ​​of Foxp3 cells⁺ CD4⁺ in the small intestine and peripheral lymph nodes, GF mice are equal to or higher than in SPF mice (see Figures 6 to 8). In addition, the number of Treg lymphocytes in the lumen of the small intestine, Peyer's patches and mesenteric lymph nodes of SPF mice that were orally administered with antibiotics for eight weeks was equal to or higher than in SPF mice (see Fig. 9). At the same time, the number of Foxp3 cells⁺CD4⁺ in the lamina propria of the large intestine, GF mice decreased significantly compared to SPF mice (see Figures 6 and 7). This decrease was commonly observed in mice with different genetic background (Balb / c, IQI and C57BL / 6) and among mice cultured in different animal facilities (see Figure 7 for data on different genetic background types, data on mice cultured in different sites for animals, no

-31 are shown in the drawings). In addition, it was also shown that the number of Treg lymphocytes in the lamina propria of SPF C57BL / 6 mice treated with antibiotics significantly decreased (see Fig. 9).

(Example 3) [0116] It was then directly verified whether the reduction in the number of Treg lymphocytes in the GF mouse luminal lamina described in Example 2 is attributed to a lack of microbiota. Specifically, a stool suspension from B6 SPF mice purchased from The Jackson Laboratory was orally administered to GF IQI mice (convention). Three weeks after the administration, lymphocytes were separated from the lamina propria of the large intestine and the expression Foxp3 was analyzed in CD4 lymphocytes⁺. Figures 10 and 11 show the results obtained. It should be remembered that each white circle in Fig. 11 shows the absolute value of CD4 cells⁺ Foxp3⁺ in each mouse separately, error bars are standard deviations (SD), * means "P &lt;0.01" in Student's t test and ** means "P &lt;0.001". As can be seen from the results shown in Figures 10 and 11, the number of Treg lymphocytes in the lamina propria of the small intestine has not changed. However, the number of Treg lymphocytes in the lamina propria of the large intestine has increased significantly. It has been shown that the host-microbial interaction played an important role in the accumulation of Treg Foxp3 lymphocytes⁺ in the lamina propria of the large intestine, and the accumulation of Treg lymphocytes in the lamina propria of the small intestine had a different mechanism.

(Example 4) [0117] Next, the relationship between lymphoid tissues bound to the gut of mice and the number of Foxp3 cells was examined.⁺ in the lamina propria of the large intestine of mice according to the method described in MN Kweon et al., J Immunol 174, 4365 (Apr. 1, 2005). Specifically, 100 μg of the recombinant extracellular domain protein (fusion protein (LTeR-Ig) between β-lymphotoxin (LTeR) and the human IgG1 Fc region, see Honda et al., J Exp Med 193, 621 (Mar 5, 2001)) were injected intraperitoneally pregnant C57BL / 6 mice 14 days after fertilization. LTeR-Ig was injected intraperitoneally with fetuses obtained from mice so that mice in which the isolated lymphoid follicles (ILF), Peyer's patches (PP) and tufts of the large intestine (CP) were completely removed. The ratio of Foxp3 cells was then analyzed⁺ in CD4 cells⁺ in the lamina propria of the large intestine of LTeR-Ig-treated mice and mice treated with rat IgG (control) using FACS. Fig. 12 shows the results obtained. It should be noted that in Fig. 12, each white circle indicates the ratio of Foxp3 cells⁺ each mouse separately, and the error bars are standard deviations. As can be seen from the results presented in Fig. 12, it was found that the ratio of Foxp3 cells⁺ in the lamina propria of the large intestine in mice deficient in isolated lymphoid vesicles, Peyer's patches and tufts of the large intestine (mice treated with LteR-Ig) increased rather. Accordingly, it has been suggested that the decrease in the number of Treg lymphocytes in the lamina propria of GF mice and mice treated with antibiotics is rather due to the fact that there is no transmission of individual signals that affect the accumulation of Treg lymphocytes in the lamella

-The proper colon, which is caused by intestinal microorganisms, and not a secondary effect of disorganized lymphoid tissues associated with the intestines.

(Example 5) [0118] To check if accumulation of colorectal lymphocytes induced a specific intestinal flora, vancomycin was administered as an Gram-positive antibiotic or polymyxin B as an antibiotic against gram-negative bacteria in SPF mice (after 4 weeks of age) for four weeks. , the ratio of Foxp3 cells was analyzed⁺in the CD4 cell group⁺ ([%] Foxp3⁺ in CD4). Fig. 30 shows the results obtained. It should be noted that in Fig. 30, "SPF" means the result in SPF mice (control), "poly B" is the result in SPY treated polymyxine B, and "Vanco" is the result obtained in SPV mice given vancomycin. By contrast, * means "P &lt;0.01".

[0119] As can be seen from the results shown in Fig. 30, the number of Treg cells in the large intestine of mice treated with vancomycin significantly decreased relative to the control. However, there was no effect on the number of Treg lymphocytes in mice treated with polymyxin B. These facts suggest that Gram-positive commensal bacteria play a major role in Treg cell accumulation.

(Example 6) [0120] A recent report suggests that spore-producing bacteria play an important role in the response of intestinal T-lymphocytes (see V. Gaboriau-Routhiau et al., Immunity 31, 677 (Oct. 16, 2009)). Accordingly, faecal microorganisms (the fraction producing spore forms) resistant to 3% chloroform were orally administered to GF mice in which the ratio of Foxp3 cells was analyzed.⁺in the CD4 cell group⁺ ([%] Foxp3⁺ in CD4). Fig. 31 shows the results obtained. It should be noted that in Fig. 31 "GF" means the result of GF and "+ chloro" mice is the result of GF mice treated with chloroform feces. However, ** means that "P &lt;0.001".

[0121] As can be seen from the results shown in Fig. 31, three weeks after faecal administration of chloroform-treated faeces, the number of Treg cells in the mouse that received it increased significantly to the same level as SPF mice and GF mice that were treated with chloroform. gavage was given by gavage (see Figures 7 and 11).

[0122] Accordingly, taking into account the results set forth in Example 5, it was found that highly probable specific components of the local microbiota belonged to the group of gram-positive bacteria and that the spore-forming fraction played an important role in the induction of Treg lymphocytes.

(Example 7) [0123] Further, intestinal microbial species were identified that induced Treg lymphocyte accumulation in the large intestine as proposed in examples 4 to 6. Specifically, segmented filamentous bacteria (SFB), 16 strains of Bacteroides spp. (Bactero. 6 strains of B. vulgatus, 7 B. acidifaciens groups 1 and 3. B. acidifaciens group 2)), 3

Lactobacillus (Lacto. L. (L. acidophilus, L. fermentum and L. murinum)) and 46 strains of Clostridium spp. (Clost. See "Itoh, K., and Mitsuoka, T. Characterization of clostridia isolated from faeces of limited flora mice and their effect on caecal size when associated with germ-free mice. Lab. Animals 19: 111-118 (1985)) ") or microbes taken from mice (SPF) cultured in a conventional environment, GF-Balb / c mice or GF IQI mice. Mice were kept in vinyl insulators for up to three weeks. CD4 cells were then isolated from the large intestine and small intestine of the mice. The number of Treg lymphocytes in the large intestine and small intestine was analyzed by flow cytometry.

[0124] Fig. 13 shows a FACS point diagram made when CD4 cells were gated⁺ in Balb / c mice. Fig. 14 shows the ratio of Foxp3 cells⁺ in CD4 cells⁺ each mouse.

[0125] It should be noted that bacteria belonging to the genus Clostridium are classified as follows by sequencing the 16S rRNA gene. Specifically, the bacterial 16S rRNA genes were amplified by PCR using a specific primer RNA pair of the 16S rRNA gene: 5'-AGAGTTTGATCMTGGCTCAG-3 '(SEQ ID NO: 19) and 5'ATTACCGCGGCKGCTG-3' (SEQ ID NO: 20) (see T Aebischer et al., Vaccination against Helicobacter pylori-induced alterations of the gastric flora in mice. FEMS Immunol. Med. Microbiol. 46, 221-229 (2006)). The 1.5-kb PCR product was then inserted into the pCR-Blunt vector. The inserts were sequenced and lined using the ClustalW computer program. The resulting 16S rRNA gene sequences of strains 1-41 from 46 Clostridium spp. Strains are set forth in SEQ ID NOs: 21-61.

[0126] As can be seen from the results shown in Figures 13 and 14, there was no effect on the number of Treg lymphocytes in the large intestine of GF mice in which filamentous bacteria (SFB) segments were colonized (see Fig. 14). In addition, mice that were colonized with a mixture of three Lactobacillus strains obtained similar results (see Fig. 14). On the other hand, it has been shown that the accumulation of Foxp3 cells⁺ in the lamina propria of the large intestine was strongly induced in mice in which 46 strains of Clostridium spp were colonized. More importantly, accumulation took place regardless of the genetic background of mice and led to an increase in numbers similar to SPF mice, despite colonization of only one type of intestinal microbiota. It was also found that colonization of Clostridium does not change the number of Treg lymphocytes in the lamina propria of the small intestine (see Fig. 14). It should be noted that when 16 strains of Bactericides spp were colonized, the number of Treg lymphocytes in the large intestine was significantly increased. However, the degree of growth varied depending on the genetic background of the mice in which the bacteria were colonized (see Figures 13 and 14).

(Example 8) [0127] Subsequently, CD4 expression, Foxp3 expression and Helios expression of LP thymic lymphocytes and SPF mouse colon, GF mouse, mice colonized with Lactobacillus and were analyzed.

Clostridium was colonized using flow cytometry. Fig. 32 and 33 show the results obtained. It should be noted that in Figs. 32 and 33, "GF" or "germ-free" means the results of GF mice, "SPF" means the results of SPF mice, "Lacto." Means the results of mice that were colonized with Lactobacillus and "Clost." means the results of mice in which Clostridium was colonized. In Fig. 32, the vertical axis shows the quantitative ratio of Helios cells^- in the Foxp3 cell group⁺ ([%] Helios^- in Foxp3⁺) and ** means that "P &lt;0.001".

[0128] As can be seen from the results shown in Figures 32 and 33, the majority of FoxP3 cells⁺ found in SPF mice or colonized Clostridium mice does not express

Helios. It should be noted that the Helios transcription factor is expressed in Treg naturally occurring thymus lymphocytes (see AM Thornton et al., J Immunol 184, 3433 (Apr. 1, 2010)). Accordingly, it has been suggested that the majority of Treg lymphocytes in SPF mice and Clostridium colonized mice are Tregs induced in the periphery, i.e. the so-called iTreg lymphocytes.

(Example 9) [0129] Next, it was investigated whether colonization of Clostridium or the like had an effect on other T cells. Specifically, SFB, 16 strains of Bacteroides spp. (Bactero.), 46 strains of Clostridium spp. (Clost.) Or microorganisms from mice cultured in a conventional environment (SPF) in GF IQI mice. Three weeks later, specific lymphocytes of the mouse large intestine were isolated and stimulated with PMA (50 ng / ml) and ionomycin (1 μg / ml) for four hours in the presence of Golgistop (BD Bioscience). Following stimulation, intracellular cytokines were stained using anti-IL-17 antibodies PE (TC11-18H10) and antibodies against IFN-g FITC (BD Bioscience) according to the instructions of the Cytofix / Cytoperm kit (BD Bioscience). Next, the quantitative ratio of IFN-γ + cells or IL-17 + cells in CD4 + cells was analyzed by flow cytometry. Figures 15 and 16 show the results obtained. It should be noted that in Figs. 15 and 16, each white circle represents the absolute value of CD4 + IFN-y + cells or the absolute value of CD4 cells⁺ IL-17⁺ each mouse separately, and error bars are standard deviations (SD). As can be seen from the results shown in Figures 15 and 16, colonization of Clostridium has no effect on Th1 cells (CD4 + IFN-γ + cells) in the large intestine, causing only a small increase in Th17 cells (CD4 cells).⁺ IL-17⁺). Accordingly, it was suggested that the type of Clostridium was a type of bacterium that specifically induced Treg lymphocytes.

(Example 10) [0130] 46 strains of Clostridium spp. Have been reported to have an effect on the accumulation of CD8 lymphocytes⁺ intestinal intraepithelial duct (IEL) in the large intestine. Accordingly, Clostridium appears to regulate the immune system in various aspects, and Clostridium exhibits considerable ability to induce and maintain Treg lymphocyte especially in the large intestine as described herein. In addition, it is known that the type of cytokine, factor

-Transforming β (TGF-β), plays an important role in the regulation of Treg lymphocyte production.

[0131] Accordingly, it was investigated whether colonization of the large intestine Clostridium provides a TGF-β-rich environment. Specifically, on the one hand, whole colons of GF mice, colonized mice of Clostridium and colonized Lactobacillus mice were cultured for 24 hours and their culture supernatants measured for active TGF-β concentrations (TGF-βΟ ELISA (the number of mice tested was four per group). Fig 34 shows the results obtained It should be noted that in Fig. 34 "GF" is the result of GF mice, "Clost." Is the result of mice colonized with Clostridium and "Lacto." Is the result of colonized Lactobacillus mice. "P &lt;0.02" and ** means "P &lt;0.001".

[0132] As can be seen from the results depicted in Fig. 34, the amount of TGF-β produced in the large intestine of mice colonized with Clostridium was significantly greater than in GF mice and mice colonized with Lactobacillus.

[0133] Next, 24 hour intestinal epithelial cells (IEC) of GF mice and mice colonized with Clostridium were cultured, and the culture supernatants were measured for active concentration of TGF-β (TGF-βΟ by ELISA (the number of mice tested was four in each group). Figure 35 shows the results obtained It should be noted that in Figure 35 "GF" means the result of GF and "Clost." Is the result of colonized Clostridium mice, while ** means "P &lt;0.001".

[0134] As can be seen from the results shown in Fig. 35, TGF-β was detected in a cultured supernatant of IEC isolated from colonized mice of Clostridium, while no TGF-β was detected in the culture supernatant of the IEC isolated from GF mice.

[0135] Subsequently, CD4 T-lymphocytes were cultured as described above⁺ spleen for five days with 50% conditioned medium in which the IEC was isolated from GF mice or colonized mice with anti-CD3 antibody in the presence or absence of an anti-TGF-β antibody. T-lymphocytes were then collected and analyzed for Foxp3 expression using real-time RT-PCR. Fig. 36 shows the results obtained. It should be noted that in Figure 36, "Medium" means the result of the medium in which the bacteria were not cultured, "GF" means the result of conditioned culture medium in which the IEC mice of GF mice were grown, "Clost." Means the result of conditioned culture medium on which the IEC mice colonized with Clostridium and "Clost. + αTGFβ "means the result of conditioned culture media into which TGF-β antibodies were added and in which IEC mice of colonized Clostridium were cultured.

[0136] As can be seen from the results shown in Fig. 36, when an IEC culture supernatant derived from Clostridium mice colonized into CD4 cells was added.⁺spleen, differentiation to Foxp3 expression was accelerated. Meanwhile, the differentiation into Treg lymphocytes was inhibited with anti-TGF-β antibodies.

[0137] In addition, the expression of MMP2, MMP9 and MMP13 was investigated, which is considered to be the reason for activation of latent TGF-β. The expression of indoleamine 2,3-dioxygenase (IDO) has also been studied, which is believed to be involved in the induction of Treg lymphocytes. Specifically, 46 strains of bacteria of the genus Clostridium (Clost.) Or three strains of bacteria of the genus Lactobacillus (Lacto.) Were administered orally to C57BL / 6 mice free of microbes. Three weeks after the administration, the IEC was collected and analyzed for mRNA expression levels of the MMP2, MMP9, MMP13 and IDO genes using real-time RTPCR (the number of mice analyzed was three per group). Figures 37 to 40 show the results obtained. It should be noted that in Figs. 37 to 40, &amp; quot; GF &amp; quot; 1 to 3 &amp; quot; are the results of GF mice, &amp; quot; Clost. # 1 to 3 &amp; quot; are the results of mice colonized with Clostridium and &amp; quot; Lacto.

[0138] For the relationship between activation of latent TGF-β and the MMP data described, see D'Angelo et al., J. Biol. Chem. 276, 11347-11353, 2001; Heidinger et al., Biol. Chem. 387, 69-78, 2006; Yu et al., Genes Dev. 14, 163-176, 2000. The relationship between IDO and Treg lymphocyte induction, see G. Matteoli et al., Gut 59, 595 (May, 2010).

[0139] As can be seen from the results shown in Figures 37 to 39, according to the described method for producing TGF-β, transcription products of genes encoding MMP2, MMP9 and MMP13 were expressed in IEC at higher levels in mice colonized with Clostridium than in GF mice and mice colonized with Lactobacillus .

[0140] In addition, as is evident from the results shown in Fig. 40, the IDO was expressed only in Clostridium colonized mice.

[0141] Accordingly, it was found that Clostridium activated IEC and led to the production of TGF-β and other molecules inducing Treg lymphocytes in the large intestine.

(Example 11) [0142] Next, it was investigated whether the accumulation of Clostridium-induced lymphocytes by Treg colonization is dependent on signal transduction through recognition receptors of molecular patterns associated with pathogens. Specifically, the number of Treg lymphocytes in the lamina propria of each SPF Myd88 mice was examined^{- / -} (deficient in Myd88 (signaling adapter for a Toll-like receptor)), Rip2^{- / -} (deficient in Rip2 (NOD receptor adapter)) and Card9^{- / -} (with Card9 deficiency (an important signal transmission factor for Dectin-1 signal transmission)). In addition, Clostridium spp. Was colonized in GF Myd88 mice^{- / -} and the change in the number of Treg lymphocytes was examined. Figures 17 and 18 show the results obtained. As can be seen from the results shown in Figs. 17 and 18, the number of Treg lymphocytes in any type of SPF mice deficient in related pathogen recognition molecules recognition receptors did not change with the wild litter mice that served as controls. In addition, it was also found that when Clostridium spp. Was colonized in GF mice deficient in Myd88, the Treg lymphocyte accumulation was induced in the lamina propria. Correspondingly, it has been suggested that the mechanism of induction of Treg lymphocyte accumulation in the lamina propria of the large intestine does not depend on the way to activate pattern recognition

-37molecular associated with major pathogens, as is the case with most bacteria, but with specific commensal bacterial species.

(Example 12) [0143] Foxp3 lymphocytes are known to be known⁺ The gastrointestinal tract performs certain immunosuppressive functions by forming IL-10 (see Non-Patent Document 9). At the same time, it is known that animals with CD4 cells⁺ Foxp3⁺those in which IL-10 has been removed in particular have developed inflammatory bowel disease (see Non-Patent Document 18). Accordingly, firstly, the expression of IL-10 on lymphocytes of various tissues was examined. In particular, lymphocytes from various tissues of SP10 Il10 mice were isolated^venus and the expression of CD4 and Venus expression were analyzed using flow cytometry. Fig. 19 shows the results obtained. It should be noted that each numerical value in Fig. 19 shows the numerical ratio of cells in the region corresponding to one of the regions divided into four.

[0144] In addition, lymphocytes of the lymphum competent mouse Il10 mice were isolated^venus and expression of the T cell e-chain receptor (TCRe) was detected on the cell surfaces using FACS. Fig. 20 shows the results obtained (FACS-spot diagrams obtained during gating of CD4 cells⁺). It is to be remembered that each numerical value in Fig. 20 shows the ratio of cells in a region corresponding to one of the regions divided into four.

[0145] In addition, lymphocytes of the lymphum competent mouse Il10 mice were isolated^venus. The lymphocytes were stimulated with PMA (50 ng / ml) and ionomycin (1 μg / ml) for four hours in the presence of Golgistop (BD Bioscience). Subsequently, after stimulation, intracellular cytokines were stained using anti-IL-17 PE antibodies, anti-IL-4 APC antibodies (11B11) and anti-IFN-g FITC antibodies (BD Bioscience) according to the instructions of the Cytofix / Cytoperm kit (BD Bioscience). Fig. 21 shows the results obtained (FACS dot diagrams obtained when placing a gateway on CD4 cells⁺). It should be remembered that each numerical value in Fig. 21 shows the ratio of cells in a region corresponding to one of the four divided regions.

[0146] In addition, Foxp3 cells were isolated⁺ CD4⁺ and Foxp3 cells^- CD4⁺ spleen (SPL) Foxp3 reporter mouse^eGFP and Venus cells were isolated⁺ from the lamina propria of the large intestine and the lamina propria of the small intestine (SI) of Il10 mice^venus . The resulting cells were then analyzed for the expression of predetermined genes. Gene expression was analyzed by RT-PCR in real time using SYBR Green PCR Master Mix (Applied Biosystems) and ABI 7300 real time PCR system (Applied Biosystems). In this case, the value for each cell was normalized to the amount of GAPDH. Fig. 22 shows the results obtained. It should be noted that in Fig. 22 the error bars are standard deviations.

[0147] As can be seen from the results shown in Figs. 19 to 22, Venus cells were hardly detected⁺ (IL10 producing cells) in the cervical lymph nodes (peripheral lymph nodes), thymus, peripheral blood, lungs and liver of mice housed

In SPF conditions. Meanwhile, a small number of Venus cells were detected in the spleen, Peyer's patches and mesenteric lymph nodes⁺ (see Fig. 19). On the other hand, many Venus cells were found⁺ in lymphocytes in the lamina propria of the small intestine and the lamina propria of the large intestine. In addition, most Venus cells⁺ in the intestines was positive CD4 cells as well as positive for the β-receptor of T cells (TCRβ) (see Figures 19 and 20). In addition, it was found that Venus + CD4 + T lymphocytes showed expression of Foxp3 and other factors associated with Treg lymphocytes, such as cytotoxic T lymphocyte antigen (CTLA-4) and glucocorticoid-induced TNFR-associated proteins (GITR), even though Venus T lymphocytes.⁺ CD4⁺ they did not show any of the Th2 phenotypes (IL-4 producing) and Th17 (IL-17 producing) phenotypes (see Figs. 21 and 22). In addition, the level of CTLA-4 expression in intestinal Venus cells was demonstrated⁺ it was higher than in Treg GFP lymphocytes⁺ spleen isolated in Foxp3 reporter mice^eGFP (see Fig. 22).

(Example 13) [0148] Venus cells⁺ can be divided into at least two subsets of cells, namely the double-positive Venus T-lymphocytes⁺ Foxp3⁺ (DP) and Treg Venus lymphocytes⁺Foxp3^- based on Foxp3 intracellular expression. The cells of the second subset correspond to Type 1 regulatory T lymphocytes (TR1) (see Non-Patent Documents 8 and 9). Accordingly, Venus cells were examined⁺ (producing IL10 cells), observed in Example 8, for the expression of Foxp3. Specifically, the expression of CD4, Foxp3 and Venus on the lamina propria of the large intestine and on the lamina propria of the small intestine of the Il10 mouse were analyzed.^venus kept under GF or SPF conditions using FACS and compared the number of Venus + cells in the lamina propria of Il10 mice^venus SPF and GF. Fig. 23 shows the results obtained (dot diagrams were obtained when CD4 cells were gated⁺).

[0149] In addition, intracellular expression of Venus and Foxp3 was analyzed in CD4 cells of various Il10 mouse tissues.^venus SPF using flow cytometry. Fig. 24 shows the results obtained (dot diagrams were obtained when the gates were placed on CD4 cells⁺). It is to be remembered that each numerical value in Fig. 24 shows the ratio of cells in a region corresponding to one of the regions divided into four.

[0150] Additionally, to determine whether the presence of commensal bacteria had an effect on the expression of IL-10 in regulatory cells in the gastro-intestinal tract, Il10 mice were prepared^venus free of microbes (GF). Then, predetermined bacterial species were colonized in the obtained Il10 mice^venus GF. Three weeks after the colonization of bacterial species, a group of CD4 cells was analyzed⁺ (V cells⁺F^-, Venus⁺ Foxp3; V cells⁺F⁺, Venus⁺ Foxp3⁺; and V cells^-F⁺, Venus^-Foxp3⁺) in which expression of Foxp3 and / or Venus was shown in the large intestine and small intestine using flow cytometry. Fig. 25 shows dot diagrams obtained when CD4 cells were gated⁺ of the large intestine, and Figures 26 and 27 show the quantitative ratio in the group of CD4 cells⁺ for each mouse. It should be remembered that each numerical value in Fig.

-3925 shows the quantitative ratio of cells in the region corresponding to one of the regions divided into four. In contrast, the error bars in Figs. 26 and 27 show standard deviations, * means "P &lt;0.02" and ** means "P &lt;0.001".

[0151] In addition, to check whether the presence of commensal bacteria had an effect on the expression of IL-10 in regulating cells in the gastrointestinal tract, antibiotics with water were given to five or six mice Il10^venus per group for 10 weeks.

The combination uses the following antibiotics.

ampicillin (A; 500 mg / l, Sigma) vancomycin (V, 500 mg / l, NACALAI TESQUE, Inc.) metronidazole (M; 1 g / l, NACALAI TESQUE, Inc.) neomycin (N; 1 g / l, NACALAI TESQUE, Inc.).

[0152] Subsequently, CD4 and Foxp3 of lymphocytes in the lumen of the large intestine, the lamina propria of the small intestine (SI), mesenteric lymph nodes (MLN) and Peyer's patches (PPS) were stained with antibodies and analyzed using FACS. The results were obtained in two or more independent experiments that gave similar results. Fig. 28 shows the results obtained (the ratio of Venus cells⁺ in CD4 cells⁺ in every attempt). It should be remembered that each white circle in Fig. 28 represents a single sample, each horizontal bar represents an average value, * indicates that "P &lt;0.02" and "AVMN" means the types of antibiotics administered under the first antibiotic letter.

[0153] As can be seen from the results shown in Figs. 23 and 24, it was found that the lamina propria of the small intestine was rich in Venus cells⁺ Foxp3^-, namely, Trl-like cells and Treg Venus lymphocytes⁺ Foxp3⁺ DPs are present at high frequency in the large intestine of SPF mice (see Figures 23 and 24). However, despite the fact that sufficient Foxp3 cells were observed⁺ also in other tissues, no Venus expression was observed in almost all cells (see Fig. 24).

[0154] In addition, as results from Figures 23 and 25 to 28, it is shown that all Venus T regulatory fractions⁺ Foxp3^-, Venus⁺ Foxp3⁺ and VenusFoxp3⁺ in the large intestine significantly decreased in GF conditions (Figures 23 and 26 to 27). In addition, a similar decrease was observed with Venus cells⁺ also in SP10 SP10 mice^venus treated with antibiotics (see Fig. 28).

[0155] In addition, as shown by the results shown in Figures 25 to 27, the colonization of Clostridium spp. Strongly induced all Venus T regulatory fractions⁺Foxp3^-, Venus⁺ Foxp3⁺ and Venus^- Foxp3⁺ in the large intestine, and the degree of induction was the same as in SPF mice (see Figs. 25 and 27). Additionally, it was found that colonization of three Lactobacillus strains or SFB colonization had very little effect on the number of Venus cells⁺ and / or Foxp3 cells⁺ in the large intestine (see Figs. 25 and 27). In addition, the colonization of 16 strains of Bacteroides spp. Also induced Venus cells⁺, but the effect of colonization was specific to cells similar to Trl Venus⁺ Foxp3^- (see Figs. 25 and 27). On the other hand

It was found that none of the bacterial species tested had any significant effect on the number of IL-10 cells produced in the lamina propria of the small intestine (see Fig. 26).

[0156] Accordingly, colonization of the Clostridium genus in the large intestine or physiologically active substances of the bacterium has been shown to provide a signal to induce accumulation of IL10 regulatory T-cells⁺ in the lamina propria of the large intestine or expression of IL-10 T-lymphocytes. At the same time, it was found that the number of Venus cells⁺ in the small intestine has not been significantly altered when commensal bacteria were not present or commensal bacteria were reduced (see Figures 23 and 26 to 28) and that IL10 regulatory lymphocytes⁺ (TR1-like cells) accumulated in the lamina propria of the small intestine independent of commensal bacteria.

(Example 14) [0157] Venus cells were tested⁺ induced by the genus Clostridium had an immunosuppressive function similar to Venus cells⁺ in the large intestine of SPF mice. Specifically, CD4 cells⁺ CD25^- (effector T cells, Teff cells) isolated from the spleen, plated on a 2 x 10 flat plate 96-well plate⁴/ well and cultured for three days, with 2 x 10⁴ CD11c cells⁺ spleen (antigen cells) followed by 30 Gy irradiation, 0.5 μg / ml anti-CD3 antibody and multiple Treg lymphocytes. In addition, CD4 + CD25 cells were grown in the last six hours^- adding [³H] -thymidine (1 μCi / well). It should be noted that the Treg lymphocytes used in Example 14 were CD4 T lymphocytes⁺ GFP⁺isolated from the spleen of Foxp3 reporter mouse mice^eGFP or CD4 T-lymphocytes⁺Venus + in the lamina propria of the GF Il10 mouse^venusin which Clostridium spp. or Il10 mice were colonized^venus SPF. Cell proliferation was then determined based on assimilation values ​​[³H] thymidine, presented in the form of counts per minute (cpm).

[0158] As can be seen from the results shown in Fig. 29, Venus cells⁺ CD4⁺ mice, in which the genus Clostridium was colonized, inhibited the in vitro proliferation of CD25-activated T lymphocytes^- CD4⁺. The inhibitory activity was slightly lower than the GFP cells⁺ isolated from Foxp3 reporter mice^eGFPbut equal to Venus cells⁺isolated from SP10 Il10 mice^venus. Accordingly, it has been shown that the type of Clostridium induces IL10 expression of T cells with a suitable immunosuppressive effect, and thus plays an important role in maintaining the immunological homeostasis of the large intestine.

(Example 15) [0159] The effect on the local immune response of colonization of a large number of Clostridium and the resulting proliferation of Treg lymphocytes was then examined.

<Model of colitis of dextran sulphate induced sodium salt (DSS)>

[0160] First, a model of DSS-induced colitis was created as described and the effect on modeled Clostridium inoculated mice and proliferation was examined.

Treg -41lymphocytes. Specifically, control mice and Clostridium inoculated mice were treated with 2% DSS, followed by weight loss, stool hardness and bleeding for six days, followed by quantification. In addition, on day 6 large intestines were collected, incised and analyzed histologically using HE staining. Figures 41 to 43 show the results obtained. It should be noted that in Figs. 41 to 43 &amp; quot; SPF + Clost. &amp; Quot; or &amp; quot; SPF + Clost. # 1 to 3 &amp; quot; are the results of C57BL / 6 mice inoculated with a colonic suspension of mice colonized with Clostridium and cultured in a conventional environment for six weeks, and SPF "or" SPF # 1 to 3 "indicate the results of C57BL / 6 mice (control mice) cultured in a conventional environment for six weeks without inoculating faecal suspension. In addition, in Fig. 41, vertical axis "disease outcome" means disease activity index (DAI) and horizontal axis "after administration of 2% DSS (d)" means the days that elapsed from the first administration of 2% DSS to mice. In addition, in Fig. 41, * indicates that "P &lt;0.02" and ** means "P &lt;0.001". At this time, Treg lymphocytes were induced by dendritic regulatory cells, playing a preventive role in the model of DSS-induced colitis (see S. Manicassamy et al., Science 329, 849 (Aug 13, 2010)).

[0161] As can be seen from the results shown in Figs. 41 to 43, the symptoms of colitis, such as weight loss and anal bleeding, were significantly inhibited in mice with high Clostridial numbers (hereinafter referred to as "mice rich in Clostridium") ) compared to control mice (see Fig. 41). All the typical features for colitis, such as intestinal shortening, edema and hemorrhage were clearly seen in control mice, as compared to mice that were clostridium-rich (see Fig. 42). In addition, histological symptoms, such as erosion of the mucous membrane, edema, cell infiltration, crypt loss, were lighter in clostridium mice treated with DSS than in control mice (see Fig. 43).

<Oxazolone-induced colitis model>

[0162] Next, an oxazolone-induced colitis model was prepared as described and the effect of Clostridium inoculation on modeled mice and Treg cell proliferation was examined. Specifically, control mice and Clostridium inoculated mice were sensitized with oxazolone followed by 1% oxazolone / 50% ethanol solution rectally. Weight loss was then observed and measured. In addition, large intestines were excised and histologically analyzed using HE staining. Fig. 44 and 45 show the results obtained. It should be noted that in Figs. 44 and 45, "SPF + Clost." Means the results of C57BL / 6 mice (Clostridium abundant mice) inoculated with a colonic suspension of Clostridium mice and grown in a conventional environment for six weeks, and "SPF" means the results of C57BL / 6 mice (control mice) cultured in a conventional environment for six weeks without inoculated faecal suspension. In addition, in Figure 44, the vertical axis "Mass (% of initial)" means the body weight after administration of 1% oxazolone, where the body weight prior to administration was taken as 100%, on the horizontal axis "after administration of 1% oxazolone (d)" means days that have elapsed since giving 1% oxazolone to mice. At the same time, it is known that Th2 type T cells contribute to the onset of inflammation

Oxalolone-induced large coli. (See M. Boirivant, IJ Fuss, A. Chu, W. Strober, J Exp Med 188, 1929 (Nov. 16, 1998)).

[0163] As results from the results shown in Figs. 44 and 45, inflammation of the large intestine with constant weight loss in control mice occurred. At the same time, the weight loss of mice abundant in Clostridium was smaller (see Fig. 44). In addition, it was also found that the symptoms of histological disease, such as erosion of the mucous membrane, edema, cell infiltration and hemorrhage, decreased in the large intestine of mice clostridium-rich (see Fig. 45).

(Example 16) [0164] Next, the effect on the systemic immune response (systemic production of IgE) of the colonization of multiple Clostridium and the resulting proliferation of Treg lymphocytes was examined. Specifically, control mice and Clostridium inoculated mice were immunized as described, giving ovalbumin to chicken (OVA) twice with an interval of 2 weeks. Serum sera were then collected and the level of OVA specific IgE was examined by ELISA. In addition, mouse spleen cells were harvested in each group and IL-4 and IL10 production was tested in vitro using OVA restimulation. Figs. 46 to 48 show the results obtained. It should be noted that in Figs. 46 to 48, "SPF + Clost." Is the results of BALB / c SPF mice (Clostridium-rich mice) inoculated with a colonic suspension of Clostridium mice colonized and cultured in a conventional environment, "SPF" means the results of BALB / c SPF mice (control mice) cultured in a conventional environment not inoculated with faecal suspension and ** means "P &lt;0.001". At the same time, in Figure 46, the vertical axis "OVA specific IgE (ng / ml)" means the concentration of serum OVA specific IgE. In addition, in Figure 46, the horizontal axis is the number of days elapsed from the first administration of ovalbumin to alum with mice rich in Clostridium or control mice (4-week-old) and "OVA + Alum" is the frequency of administration of ovalbumin with alum. In addition, in Figs. 47 and 48, &amp; quot; OVA &amp; quot; on the horizontal axis means the results in the case of OVA restimulation in vitro and "-" means the results in the case where OVA restimulation has not been performed in vitro. Additionally, in Figs. 47 and 48,

[0165] As can be seen from the results shown in Figs. 46 to 48, IgE levels were significantly lower in mice that were plentiful in Clostridium than in control mice (see Figure 46). In addition, IL-4 production by OVA restimulation decreased (see Fig. 47) and IL10 production increased (see Fig. 48) in the splenic cells of mice clustrid with OVA-coated allergens with alum compared to control mice.

[0166] Accordingly, taking into account the results given in Example 15 in total, it was found that the induction of Treg lymphocytes by Clostridium of the large intestine plays an important role in local and systemic immune responses.

(Example 17)

[0167] Subsequently, Balb / c GF mice were colonized with three strains of Clostridium cluster IV (strains 22, 23 and 32, shown in Figure 49). Three weeks later, Treg Foxp3 lymphocytes were analyzed⁺ large intestine using FACS. Fig. 50 shows the results obtained. As can be seen from the results shown in Fig. 50, gnotobiotic mice colonized with three Clostridium strains showed a Treg lymphocyte induction pattern intermediate between GF mice and mice inoculated with all strains.

(Example 18) [0168] It was then examined whether the fractions producing spore forms (e.g., chloroform resistant) human faeces samples induced the proliferation or accumulation of regulatory T lymphocytes, similar to the fraction producing the spore forms of a stool sample obtained from mice.

[0169] Specifically, human healthy volunteer (Japanese, 29 years old) faeces were suspended in phosphate buffered saline (PBS), mixed with chloroform (3% final concentration), and then incubated by shaking in a bath for 60 min. After evaporation of chloroform using N2 gas bubbling, aliquots containing fractions resistant to chloroform (e.g., spore forms) of human intestinal bacteria were inoculated orally into microbial-free (GF) mice (IQI, age 8 weeks). Treated mice were kept in a vinyl insulator for 3 weeks. The large intestine was harvested and dissected longitudinally, washed to remove stool content, and shaken in Hanks's balanced salt solution (HBSS) containing 5 mM EDTA for 20 minutes at 37 ° C. After removal of epithelial cells and adipose tissue, large intestine was cut into small pieces and incubated with RPMI1640 containing 4% fetal bovine serum, 1 mg / ml collagenase D, 0.5 mg / ml dispase and 40 μg / ml DNase I (all manufactured by Roche Diagnostics) for 1 hour at room temperature 37 ° C in a shaking bath. The digested tissue was washed with HBSS containing 5 mM EDTA, resuspended in 5 ml 40% Percoll (manufactured by GE Healthcare) and applied with 2.5 ml 80% Percoll in a 15 ml Falcon tube. Separate Percoll gradient by centrifugation at 780 g for 20 minutes at 25 ° C. The cells were harvested at the interface and suspended in staining buffer containing PBS, 2% FBS, 2 mM EDTA and 0.09% NaN3 and stained on the surface of CD4 with anti-CD4 Ab-labeled anti-CD4 (RM4-5, manufactured by BD Biosciences). Intracellular Foxp3 staining was performed using Alexa647 labeled against Foxp3 Ab (FJK-16, produced by eBioscience) and Foxp3 Staining Buffer Set (produced by eBioscience). The percentage of positive Foxp3 cells in the CD4 positive cell population was analyzed using flow cytometry. Figs. 51 and 52 show the results obtained.

[0170] The figures show histograms (Fig. 51) and combined data (Fig. 52) of Foxp3 expression on CD4 positive GF mice (GF) or GF mice injected with a human feces treated with chloroform (GF + Chloro.) . In addition, the numbers in Fig. 51 indicate the percentage of cells in the gate. Each circle in the drawing. 52 refers to a separate animal, error bars are SD and ** mean "P &lt;0.001".

[0171] As can be seen from the results shown in Figs. 51 and 52, it was also found that when (for example chloroform resistant) fractions forming spore forms of human intestinal bacteria were colonized in GF mice, the accumulation of Foxp3 regulatory lymphocytes was induced.⁺ (Treg) in the lamina propria of the large mouse colon.

[0172] Subsequently, it was examined which bacterial species increased after being administered by gavage of a human stool treated with chloroform.

[0173] Specifically, using QIAamp DNA Stool mini kit (manufactured by Qiagen), bacterial genomic DNA was isolated from the human feces of a healthy volunteer, as described (human stool) or GF mouse stool granules, which were administered by gavage to human stool treated with chloroform ( GF + Chloro). Quantitative PCR analysis was performed using LightCycler 480 (manufactured by Roche). The relative value was calculated by the method of ACt and normalized to the total amount of bacteria, dilution and weight of the sample. The following starter sets were used:

Total number of bacteria

<td>GGTGAATACGTTCCCGG 5'-3 '</td><td>(SEQ ID</td><td></td><td>NO:</td><td>62)</td><td>and</td>

<td>TACGGCTACCTTGTTACGACTT 5'-3 '</td><td>(KNOT</td><td>ID</td><td></td><td>NO:</td><td>63)</td>

<td colspan="2">Clostridium cluster XIVa (Clostridium coccoides subgroup).</td><td></td><td></td><td></td><td></td>

<td>[0174]</td><td></td><td></td><td></td><td></td><td></td>

<td>AAATGACGGTACCTGACTAA 5'-3 '</td><td colspan="2">(SEQ ID</td><td>NO:</td><td>64)</td><td>and</td>

<td colspan="2">5'-CTTTGAGTTTCATTCTTGCGAA-3 '(SEQ ID NO: 65)</td><td></td><td></td><td></td><td></td>

<td colspan="5">[0175] Clostridium cluster IV (Clostridium leptum) 5'-GCACAAGCAGTGGAGT-3 '</td><td>(KNOT</td>

<td>ID NO:</td><td></td><td>66)</td><td></td><td></td><td>and</td>

<td>CTTCCTCCGTTTTGTCAA 5'-3 ' Bacteroides</td><td>(KNOT</td><td>ID</td><td></td><td>NO:</td><td>24)</td>

<td>GAGAGGAAGGTCCCCCAC 5'-3 '</td><td>(SEQ ID</td><td></td><td>NO:</td><td>67)</td><td>and</td>

5'-CGCTACTTGGCTGGTTCAG-3 '(SEQ ID NO: 68).

[0176] Fig. 53 shows the results obtained.

[0177] As can be seen from the results shown in Fig. 53, mice injected with chloroform-treated human stools were characterized by high amounts of bacteria forming spore forms, such as Clostridium clusters XIVa and IV, and a significant decrease in non-spore bacteria, such as as Bacteroides, compared to the human stool before subjecting it to chloroform.

[Industrial application] [0178] As described, the invention enables providing a perfect composition for inducing the proliferation or accumulation of regulatory T cells (Treg cells) by using bacteria belonging to the genus Clostridium or a physiologically active substance or the like, a bacterial derivative. Since the composition of the invention has an immunosuppressive effect, the composition can be used, e.g.

- preventing or treating autoimmune diseases and allergic diseases, as well as in inhibiting immune rejection in organ transplants or the like. In addition, healthy people can easily and routinely take the composition in the form of food or beverages, such as health-promoting foods that improve immune function.

[Sequence list] [0179]

SEQ ID NO: 1 to 20, 62 to 69 <223> Artificially synthesized primer sequence [0180] SEQ ID NO: 21 to 61 <223> The sequence of the 16S rRNA gene for each Clostridium strain

Sequence list [0181] <110> The University of Tokyo <120> Composition for inducing proliferation or accumulation of regulatory T lymphocytes <130> 11B1040011 <150> JP2010 / 129134 <151> 2010-06-04 <150> PCT / JP2010 / 071746 <151> 2010-12-03 <160> 69 <170> Patent version 3.1 <210> 1 <211> 23 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 1 ggcaatagtt ccttcccaga gtt 23 <210> 2 <211> 22 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence

-46 <400> 2 gggtcgcata ttgtggtact tg 22 <210> 3 <211> 23 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 3 ccttttgtag ccctgctcac tct 23 <210> 4 <211> 21 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 4 gggtcacctg tatggcttca g 21 <210> 5 <211> 21 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 5 tcagtgcaag atctgcaagc a 21 <210> 6 <211> 19 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 6 acaccggaag ccaaacaca 19

-47 <210> 7 <211> 27 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 7 gattttaata agctccaaga ccaaggt 27 <210> 8 <211> 27 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 8 cttctatgca gttgatgaag atgtcaa 27 <210> 9 <211> 21 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 9 cctcgtcccg tagacaaaat g 21 <210> 10 <211> 20 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 10 tctccacttt gccactgcaa 20 <210> 11 <211> 20 <212> DNA

-48 <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 11 ggacattgtc tttgatggca 20 <210> 12 <211> 20 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 12 cttgtcacgt ggtgtcactg 20 <210> 13 <211> 20 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 13 tctctggacg tcaaatgtgg 20 <210> 14 <211> 19 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 14 gctgaacagc agagccttc 19 <210> 15 <211> 20 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence

-49 <400> 15 aggtctggat cactccaagg 20 <210> 16 <211> 19 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 16 tcgcctggac cataaagaa 19 <210> 17 <211> 20 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 17 agaggatgcg tgactttgtg 20 <210> 18 <211> 20 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 18 atacagcaga ccttctggca 20 <210> 19 <211> 20 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 19 agagtttgat cmtggctcag 20

-50 <210> 20 <211> 16 <212> DNA <213> Artificial <220>

<223> Artificially synthesized primer sequence <400> 20 attaccgcgg ckgctg 16 <210> 21 <211> 1460 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1460) <223> The sequence of the gene encoding 16S rRNA 1 of the Clostridium strain

-51 <400> 21

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gagaaccatt</td><td>ggatcgagga</td><td>ttcgtccaag</td><td>tgaaggtggg</td><td>gaaagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtga</td><td>gcaatctgcc</td><td>ttggagtggg</td><td>gaataacggc</td><td>tggaaacagc</td><td>cgctaatacc</td><td>180</td>

<td>gcatgataca</td><td>gctgggaggc</td><td>atctccctgg</td><td>ctgtcaaaga</td><td>tttatcgctc</td><td>tgagatgagc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggcggggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgcaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttgtc</td><td>agggacgaag</td><td>caagtgacgg</td><td>tacctgacac</td><td>480</td>

<td>ggctactacg</td><td>gtcagcagcg</td><td>cgtatacgta</td><td>ggtgccagcg</td><td>tatccggaat</td><td>tacctgggtt</td><td>540</td>

<td>aaggcgtgtt</td><td>agccggactg</td><td>cagtcagatg</td><td>tgaatcacgg</td><td>gctcaacttg</td><td>tgctgcattg</td><td>600</td>

<td>gaactgtagt</td><td>tctgagtact</td><td>gagagcagac</td><td>ggaattctag</td><td>gtagcggtga</td><td>atgcgtagat</td><td>660</td>

<td>ataggaggac</td><td>acagtgcgag</td><td>gcgtctgctg</td><td>acagcaactg</td><td>acgctgaggc</td><td>gggaagcgtg</td><td>720</td>

<td>ggggagccaa</td><td>caggattaga</td><td>tacctggtag</td><td>ttcacgcctg</td><td>gtaaaacgat</td><td>ggatactagg</td><td>780</td>

<td>tgtgggggga</td><td>ctgaccccct</td><td>cgtggccgcc</td><td>agttaacacc</td><td>aataaagtat</td><td>cccacctggg</td><td>840</td>

<td>agtacgatcg</td><td>caaggttgaa</td><td>actcaaagga</td><td>attgacgggg</td><td>cccgcacaag</td><td>cggtggagta</td><td>900</td>

<td>tgtggtttaa</td><td>ttcgaagcaa</td><td>cgcgaagaac</td><td>cttaccaggg</td><td>cttgacatcc</td><td>cgaggaccgg</td><td>960</td>

<td>actagagata</td><td>gtcttttctc</td><td>ttcggagacc</td><td>tcggtgacag</td><td>gtggtgcatg</td><td>gttgtcgtca</td><td>1020</td>

<td>gctcgtgtcg</td><td>taagatgttg</td><td>ggttaagtcc</td><td>cgcaacgagc</td><td>gcaaccctta</td><td>ttgttagttg</td><td>1080</td>

<td>ctacgcaaga</td><td>gcactctagc</td><td>gagactgccg</td><td>ttgacaaaac</td><td>ggaggaaggt</td><td>ggggacgacg</td><td>1140</td>

<td>tcaaatcatc</td><td>atgcccctta</td><td>tgtcctgggc</td><td>cacacacgta</td><td>ctacaatggt</td><td>ggtcaacaga</td><td>1200</td>

<td>gggaagcaat</td><td>accgcgaggt</td><td>ggagcaaatc</td><td>cctaaaagcc</td><td>atcccagttc</td><td>ggatcgcagg</td><td>1260</td>

<td>ctgcaacccg</td><td>cctgcgtgaa</td><td>gttggaatcg</td><td>ctagtaatcg</td><td>cggatcagca</td><td>tgccgcggtg</td><td>1320</td>

<td>aatacgttcc</td><td>cgggccttgt</td><td>acacaccgcc</td><td>cgtcacacca</td><td>tgagagtcgg</td><td>gaacacccga</td><td>1380</td>

<td>agtccgtagc</td><td>ctaaccgcaa</td><td>gggggggcgc</td><td>ggccgaaggt</td><td>gggttcgata</td><td>attggggtga</td><td>1440</td>

<td>agtcgtaaca</td><td>aggtagccgt</td><td></td><td></td><td></td><td></td><td>1460</td>

<210> 22 <211> 1485 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1485) <223> The sequence of the gene encoding 16S rRNA 2 of the Clostridium strain

-52 <400> 22

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggaagggact</td><td>gcttagtggc</td><td>ggacgggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>cttggagtgg</td><td>ggaataacag</td><td>ctggaaacag</td><td>ctgctaatac</td><td>180</td>

<td>cgcataatat</td><td>atctgggccg</td><td>catggctctg</td><td>gatatcaaag</td><td>atttatcgct</td><td>ctgagatgga</td><td>240</td>

<td>ctcgcgtctg</td><td>attagctagt</td><td>tggcggggta</td><td>acggcccacc</td><td>aaggcgacga</td><td>tcagtagccg</td><td>300</td>

<td>gactgagagg</td><td>ttggccggcc</td><td>acattgggac</td><td>tgagacacgg</td><td>cccagactcc</td><td>tacgggaggc</td><td>360</td>

<td>agcagtgggg</td><td>aatattgggc</td><td>aatgggcgca</td><td>agcctgaccc</td><td>agcaacgccg</td><td>cgtgaaggaa</td><td>420</td>

<td>gaaggctttc</td><td>gggttgtaaa</td><td>cttcttttgt</td><td>cagggacgaa</td><td>gcaagtgacg</td><td>gtacctgacg</td><td>480</td>

<td>aataagccac</td><td>ggctaactac</td><td>gtgccagcag</td><td>ccgcggtaat</td><td>acgtaggtgg</td><td>caagcgttat</td><td>540</td>

<td>ccggatttac</td><td>tgggtgtaaa</td><td>gggcgtgtag</td><td>gcgggactgc</td><td>aagtcagatg</td><td>tgaaaaccac</td><td>600</td>

<td>gggctcaacc</td><td>tgtgggcctg</td><td>catttgaaac</td><td>tgtagttctt</td><td>gagtactgga</td><td>gaggcagacg</td><td>660</td>

<td>gaattctagt</td><td>tgtagcgtga</td><td>aatgcgtaga</td><td>tatagaagaa</td><td>cacagttgcg</td><td>gagccggtct</td><td>720</td>

<td>gcaactgacg</td><td>ctgagcgcga</td><td>aagcgtgggg</td><td>agcaaacagg</td><td>attagatacc</td><td>ctggtagtcc</td><td>780</td>

<td>acgctgtaaa</td><td>cgatggatta</td><td>ctaggtgtgg</td><td>ggggactgac</td><td>cccctccgtg</td><td>ccgcagttaa</td><td>840</td>

<td>cacaataagt</td><td>atcccacctg</td><td>gggagtacga</td><td>tcgcaaggtt</td><td>gaaactcaaa</td><td>aggaattgac</td><td>900</td>

<td>gggggcccgc</td><td>acaagcggtg</td><td>gagtatgtgg</td><td>tttaaattcg</td><td>aagcaacgcg</td><td>aagaacctta</td><td>960</td>

<td>ccagggcttg</td><td>acatcccggt</td><td>gaccgtccta</td><td>gagataggat</td><td>tttcccttcg</td><td>gggacactgg</td><td>1020</td>

<td>agacaggtgg</td><td>tgcatggttg</td><td>tcgtcagctc</td><td>gtgtcgtgag</td><td>atgttgggtt</td><td>aagtcccgca</td><td>1080</td>

<td>acgagcgcaa</td><td>cccttattgt</td><td>tagttgctac</td><td>gcaagagcac</td><td>tctagcgaga</td><td>ctgccgttga</td><td>1140</td>

<td>caaaacggag</td><td>gaaggtgggg</td><td>acgacgtcaa</td><td>atcatcatgc</td><td>cccttatgtc</td><td>ctgggccaca</td><td>1200</td>

<td>cacgtactac</td><td>aatggtggtc</td><td>aacagaggga</td><td>agcaaagccg</td><td>cgaggtggag</td><td>caaatcccta</td><td>1260</td>

<td>aaagccatcc</td><td>cagttcggat</td><td>cgcaggctgc</td><td>aacccgcctg</td><td>cgtgaagttg</td><td>gaatcgctag</td><td>1320</td>

<td>taatcgcgga</td><td>tcagaatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>ccttgtacac</td><td>accgcccgtc</td><td>1380</td>

<td>acaccatgag</td><td>agtcgggaac</td><td>acccgaagtc</td><td>cgtagcctaa</td><td>ccgcaagggg</td><td>ggcgcggccg</td><td>1440</td>

<td>aaggtgggtt</td><td>cgataattgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td></td><td>1485</td>

<210> 23 <211> 1491 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1491) <223> The sequence of the gene encoding 16S rRNA 3 of the Clostridium strain

-53 <400> 23

<td>agagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacctt</td><td>cgaaagagac</td><td>ttcggtcaat</td><td>ggaaaagaat</td><td>gcttagtggc</td><td>ggacgggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>ctttcagtgg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcataacgt</td><td>acgggtatcg</td><td>catggtatct</td><td>gtaccaaaga</td><td>tttatcgctg</td><td>agagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggtagggtaa</td><td>cggcctacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgaaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttgac</td><td>ggggaagagc</td><td>agaagacggt</td><td>acctgtcgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttgtcc</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gcgtgtagcc</td><td>gggctgacaa</td><td>gtcagatgtg</td><td>aaatccgggg</td><td>600</td>

<td>gctcaacccc</td><td>cgaactgcat</td><td>ttgaaactgt</td><td>tggtcttgag</td><td>tatcggagag</td><td>gcaggcggaa</td><td>660</td>

<td>ttcctagtgt</td><td>agcggtgaaa</td><td>tgcgtagata</td><td>ttagggggaa</td><td>caccagtggc</td><td>gaagcggcct</td><td>720</td>

<td>gctggacgac</td><td>aactgacggt</td><td>gaggcgcgaa</td><td>agcgtgggga</td><td>gcaaacagga</td><td>ttagataccc</td><td>780</td>

<td>tggtagtcca</td><td>cgctgtaaac</td><td>gatggatact</td><td>aggtgtgcgg</td><td>ggactgaccc</td><td>ctgcgtgccg</td><td>840</td>

<td>cagctaacgc</td><td>aataagtatc</td><td>ccacctgggg</td><td>agtacgatcg</td><td>caaggttgaa</td><td>actcaaagga</td><td>900</td>

<td>attgacgggg</td><td>gcccgcacaa</td><td>gcggtggatt</td><td>atgtggttta</td><td>attcgatgca</td><td>acgcgaagaa</td><td>960</td>

<td>ccttaccagg</td><td>gcttgacatc</td><td>ctactaacga</td><td>agtagagata</td><td>cattaggtac</td><td>ccttcggggg</td><td>1020</td>

<td>aagtagagac</td><td>aggtggtgca</td><td>tggttgtcgt</td><td>cagctcgtgt</td><td>cgtgagatgt</td><td>tgggttaagt</td><td>1080</td>

<td>cccgcaacga</td><td>gcgcaaccct</td><td>tattgttagt</td><td>tgctacgcaa</td><td>gagcactcta.</td><td>gcgagactgc</td><td>1140</td>

<td>cgttgacaaa</td><td>acggaggaag</td><td>gtggggacga</td><td>cgtcaaatca</td><td>tcatgcccct</td><td>tatgtcctgg</td><td>1200</td>

<td>gctacacacg</td><td>taatacaatg</td><td>gcggtcaaca</td><td>gagggatgca</td><td>aaaccgcgag</td><td>gtggagcgaa</td><td>1260</td>

<td>cccctaaaag</td><td>ccgtcccagt</td><td>tcagatcgca</td><td>gtctgcaacc</td><td>cgactgcgtg</td><td>aagtcggaat</td><td>1320</td>

<td>cgctagtaat</td><td>cgcggatcag</td><td>catgccgcgg</td><td>tgaatacgtt</td><td>cccgggcctt</td><td>gtacacaccg</td><td>1380</td>

<td>cccgtcacac</td><td>catgagagtc</td><td>gggaacaccc</td><td>gaagtccgta</td><td>gcctaaccgc</td><td>aaggagggcg</td><td>1440</td>

<td>cggccgaagg</td><td>tgggttcgat</td><td>aattggggtg</td><td>aagtcgtaac</td><td>aaggtagccg</td><td>t</td><td>1491</td>

<210> 24 <211> 1491 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1) ... (1491) <223> The sequence of the gene encoding the 16S rRNA of the 4 strain Clostridium <400> 24

-54agagtttgat cctggctcag gatgaacgct ggcggcgtgc ctaacacatg caagtcgaac 60 gggtgtacgg gggagaaggc ttcggccgga aaacctgtgc atgagtggcg gacgggggg 120

<td>taacgcgtgg</td><td>gcaacctggc</td><td>ctgtacaggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>180</td>

<td>gcataacggg</td><td>ggaagccgca</td><td>tggcttttcc</td><td>ctgaaaactc</td><td>cggtggtaca</td><td>ggatgggccc</td><td>240</td>

<td>gcgtctgatt</td><td>agccagttgg</td><td>cagggtaacg</td><td>gcctaccaaa</td><td>gcgacgatca</td><td>gtagccggcc</td><td>300</td>

<td>tgagagggcg</td><td>gacggccaca</td><td>ctgggactga</td><td>gacacggccc</td><td>agactcctac</td><td>gggaggcagc</td><td>360</td>

<td>agtgggggat</td><td>attgcacaat</td><td>ggggggaaac</td><td>cctgatgcag</td><td>cgacgccgcg</td><td>tgagtgaaga</td><td>420</td>

<td>agtatttcgg</td><td>tatgtaaagc</td><td>tctatcagca</td><td>gggaagaaaa</td><td>tgacggtacc</td><td>tgactaagaa</td><td>480</td>

<td>gccccggcta</td><td>actacgtgcc</td><td>agcagccgcg</td><td>gtaatacgta</td><td>gggggcaagc</td><td>gttatccgga</td><td>540</td>

<td>tttactgggt</td><td>gtaaagggag</td><td>cgtagacggc</td><td>agcgcaagtc</td><td>tgagtgaaat</td><td>cccatggctt</td><td>600</td>

<td>aaccatggaa</td><td>ctgctttgga</td><td>aactgtgcag</td><td>ctggagtgca</td><td>ggagagtaag</td><td>cggaattcct</td><td>660</td>

<td>agtgtagcgt</td><td>gaaatgcgta</td><td>gattatagga</td><td>ggaacaccag</td><td>tggcgaaggc</td><td>ggctaactga</td><td>720</td>

<td>actgtaactg</td><td>acgttgaggc</td><td>tcgaaagcgt</td><td>ggggagcaaa</td><td>caggattaga</td><td>taccctggta</td><td>780</td>

<td>gtccacgccg</td><td>taaacgatga</td><td>ttactaggtg</td><td>ttgggggacc</td><td>aaggtcttcg</td><td>gtgccggcgc</td><td>840</td>

<td>aaacgcatta</td><td>agtaatccac</td><td>ctggggagta</td><td>cgttcgcaag</td><td>aatgaaactc</td><td>aaaggaattg</td><td>900</td>

<td>acggggaccc</td><td>gcacaagcgg</td><td>tggagcatgt</td><td>ggtttaattc</td><td>gaagcaacgc</td><td>gaagaacctt</td><td>960</td>

<td>acctggtctt</td><td>gacatcccga</td><td>tgacgagtga</td><td>gcaaagtcac</td><td>tttcccttcg</td><td>gggcattgga</td><td>1020</td>

<td>gacaggtggt</td><td>gcatggttgt</td><td>cgtcagctcg</td><td>tgtcgtgaga</td><td>tgttgggtta</td><td>agtcccgcaa</td><td>1080</td>

<td>cgagcgcaac</td><td>ccctatttcc</td><td>agtagccagc</td><td>aggtagagct</td><td>gggcactctg</td><td>gagagactgc</td><td>1140</td>

<td>ccgggataac</td><td>cgggaggaag</td><td>gcggggatga</td><td>cgtcaaatca</td><td>tcatgcccct</td><td>tatgatcagg</td><td>1200</td>

<td>gctacacacg</td><td>tgctacaatg</td><td>gcgtaaacaa</td><td>agggaagcga</td><td>gacggtgacg</td><td>ttgagcaaat</td><td>1260</td>

<td>cccaaaaata</td><td>acgtcccagt</td><td>tcggattgta</td><td>gtctgcaact</td><td>cgactacatg</td><td>aagctggaat</td><td>1320</td>

<td>cgctagtaat</td><td>cgcgaatcag</td><td>aatgtcgcgg</td><td>tgaatacgtt</td><td>cccgggtctt</td><td>gtacacaccg</td><td>1380</td>

<td>cccgtcacac</td><td>catgggagtc</td><td>ggaaatgccc</td><td>gaagtcagtg</td><td>acctaaccga</td><td>aaggaaggag</td><td>1440</td>

<td>ctgccgaagg</td><td>tggagccggt</td><td>aactggggtg</td><td>aagtcgtaac</td><td>aaggtagccg</td><td>t</td><td>1491</td>

<210> 25 <211> 1467 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1467) <223> Sequence of the gene encoding the 16S rRNA of the Clostridium strain <400> 25

<td>agagtttgat</td><td>cctggctcag gacgaacgct ggcggcacgc ctaacacatg caagtcgaac</td><td>60</td>

<td>ggagtgaaga</td><td>tgctcgcatc tgaacttagt ggcggacggg tgagtaacac gtgagcaacc</td><td>120</td>

<td>tgcctttcag</td><td>agggggatta cgtttggaaa cgaacgctaa taccgcataa aatatcggag</td><td>180</td>

<td>tcgcatggca</td><td>ctgatatcaa aggagcaatc cgctgaaaga tgggctcgcg tccgattagg</td><td>240</td>

<td>cagttggcgg</td><td>ggtatcggcc caccaaaccg acaatcggta gccggactga gaggttgaac</td><td>300</td>

<td>ggccacattg</td><td>ggactgagac</td><td>gcggccca ga</td><td>ctcctacggg</td><td>aggcagcagt</td><td>gggggatatt</td><td>360</td>

<td>gcacaatggg</td><td>ggaaaccctg</td><td>atgcagcgat</td><td>gccgcgtgaa</td><td>tgaagacggc</td><td>cttcgggttg</td><td>420</td>

<td>taaagttctg</td><td>tcgcagggga</td><td>cgaaaatgac</td><td>ggtaccctgc</td><td>aagaaagctc</td><td>cggctaacta</td><td>480</td>

<td>cgtgccagca</td><td>gccgcggtaa</td><td>tacgtaggga</td><td>gcaagcgttg</td><td>tccggaatta</td><td>ctgggtgtaa</td><td>540</td>

<td>agggagcgta</td><td>ggcgggagga</td><td>taagttgaat</td><td>gtgaaatcta</td><td>tgggctcaac</td><td>ccatagctgc</td><td>600</td>

<td>gttcaaactg</td><td>ttcttcttga</td><td>gtgaagtaga</td><td>ggcaggcgga</td><td>attcctagtg</td><td>tagcggtgaa</td><td>660</td>

<td>atgcgtagat</td><td>attaggagga</td><td>caccagtggc</td><td>gaaggcgggc</td><td>tgctgggctt</td><td>tactgacgct</td><td>720</td>

<td>gaggctcgaa</td><td>agcgtgggta</td><td>gcaaacagga</td><td>ttagataccc</td><td>tggtagtcca</td><td>cgcggtaaac</td><td>780</td>

<td>gatgattact</td><td>aggtgtgggt</td><td>ggactgaccc</td><td>catccgtgcc</td><td>ggagttaaca</td><td>caataagtaa</td><td>840</td>

<td>tccacctggg</td><td>gagtacggcc</td><td>gcaaggttga</td><td>aactcaaagg</td><td>aattgacggg</td><td>ggcccgcaca</td><td>900</td>

<td>agcagtggag</td><td>tatgtggttt</td><td>aattcgacgc</td><td>aacgcgaaga</td><td>accttaccag</td><td>gtcttgacat</td><td>960</td>

<td>cgagtgacgg</td><td>acatagagat</td><td>atgtctttcc</td><td>ttcgggacac</td><td>gaagacaggt</td><td>ggtgcatggt</td><td>1020</td>

<td>tgtcgtcagc</td><td>tcgtgtcgtg</td><td>agatgttggg</td><td>ttaagtcccg</td><td>caacgagcgc</td><td>aacccttacc</td><td>1080</td>

<td>attagttgct</td><td>acgcaagagc</td><td>actctaatgg</td><td>gactgccgtt</td><td>gacaaaacgg</td><td>aggaaggtgg</td><td>1140</td>

<td>ggatgacgtc</td><td>aaatcatcat</td><td>gccccttatg</td><td>acctgggcga</td><td>cacacgtact</td><td>acaatggcgg</td><td>1200</td>

<td>tcaacagagg</td><td>gaggcaaagc</td><td>cgcgaggcag</td><td>agcaaacccc</td><td>taaaagccgt</td><td>ctcagttcgg</td><td>1260</td>

<td>attgcaggct</td><td>gcaactcgcc</td><td>tgcatgaagt</td><td>cggaattgct</td><td>agtaatcgcg</td><td>gatcagcatg</td><td>1320</td>

<td>ccgcggtgaa</td><td>tacgttcccg</td><td>ggccttgtac</td><td>acaccgcccg</td><td>tcacaccatg</td><td>agagccggta</td><td>1380</td>

<td>acacccgaag</td><td>tcaatagtct</td><td>aaccgcaagg</td><td>aggacattgc</td><td>cgaaggtggg</td><td>attggtaatt</td><td>1440</td>

<td>ggggtgaagt</td><td>cgtaacaagg</td><td>tagccgt</td><td></td><td></td><td></td><td>1467</td>

<210> 26 <211> 1474 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1) ... (1474) <223> The sequence of the gene encoding the 16S rRNA of the 6 strain Clostridium <400> 26

<td>agagtttgat</td><td>catggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggtgtacgg</td><td>gaaggaaggc</td><td>ttcggccgga</td><td>aaacctgtgc</td><td>atgagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtgg</td><td>gcaacctggc</td><td>ctgtacaggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>180</td>

<td>gcataacggg</td><td>ggaagccgca</td><td>tggcttttcc</td><td>ctgaaaactc</td><td>cggtggtaca</td><td>ggatgggccc</td><td>240</td>

<td>gcgtctgatt</td><td>agccagttgg</td><td>cagggtaacg</td><td>gcctaccaaa</td><td>gcgacgatca</td><td>gtagccggcc</td><td>300</td>

<td>tgagagggcg</td><td>gacggccaca</td><td>ctgggactga</td><td>gacacggccc</td><td>agactcctac</td><td>gggaggcagc</td><td>360</td>

<td>agtgggggat</td><td>attgcacaat</td><td>ggggggaacc</td><td>ctgatgcagc</td><td>gacgccgcgt</td><td>gggtgaagaa</td><td>420</td>

<td>gcgcctcggc</td><td>gcgtaaagcc</td><td>ctgtcagcag</td><td>ggaagaaaat</td><td>gacggtacct</td><td>gaagaagaag</td><td>480</td>

<td>ccccggctaa</td><td>ctacgtgcca</td><td>gcagccgcgg</td><td>taatacgtag</td><td>gggggcaagc</td><td>gttatccgga</td><td>540</td>

<td>tttactgggt</td><td>gtaaaggggg</td><td>cgcagacggc</td><td>gatgcaagcc</td><td>aggagtgaaa</td><td>gcccggggcc</td><td>600</td>

<td>caaccccggg</td><td>actgctcttg</td><td>ggaactgcgt</td><td>ggctggagtg</td><td>cagagggcag</td><td>cggaattcct</td><td>660</td>

<td>ggtgaaatgc</td><td>gtagatatca</td><td>gaagacacgg</td><td>tgcgaggcgg</td><td>cctgctgact</td><td>gcactgacgt</td><td>720</td>

<td>tgagccgaag</td><td>cgtggggagc</td><td>aaacaggatt</td><td>agataccgtg</td><td>gtagtcacgc</td><td>cgtaaacgat</td><td>780</td>

<td>gattactagg</td><td>tgtcggggag</td><td>cagagactgc</td><td>ccggtgccgc</td><td>agccaacgca</td><td>ttaagtaatc</td><td>840</td>

<td>cacctgggga</td><td>gtacgttcgc</td><td>aagaatgaaa</td><td>ctcaaaggaa</td><td>ttgacgggga</td><td>cccgcacaag</td><td>900</td>

<td>cggtggagca</td><td>tgtggtttaa</td><td>ttcgaagcaa</td><td>cgcgaagaac</td><td>cttacctggt</td><td>cttgacatcc</td><td>960</td>

<td>cgatgacgag</td><td>tgagcaaagt</td><td>cactttccct</td><td>tcggggcatt</td><td>ggagacaggt</td><td>ggtgcatggt</td><td>1020</td>

<td>tgtcgtcagc</td><td>tcgtgtcgtg</td><td>agatgttggg</td><td>ttaagtcccg</td><td>caacgagcgc</td><td>aacccctatt</td><td>1080</td>

<td>tccagtagcc</td><td>agcaggtaga</td><td>gctgggcact</td><td>ctggagagac</td><td>tgcccgggat</td><td>aaccgggagg</td><td>1140</td>

<td>aaggcgggga</td><td>tgacgtcaaa</td><td>tcatcatgcc</td><td>ccttatgatc</td><td>agggctacac</td><td>acgtgctaca</td><td>1200</td>

<td>atggcgtaaa</td><td>caaagggaag</td><td>cgagacggtg</td><td>acgttaagca</td><td>aatcccaaaa</td><td>ataacgtccc</td><td>1260</td>

<td>agttcggatt</td><td>gtagtctgca</td><td>actcgactac</td><td>atgaagctgg</td><td>aatcgctagt</td><td>aatcgcgaat</td><td>1320</td>

<td>cagaatgtcg</td><td>cggtgaatac</td><td>gttcccgggt</td><td>cttgtacaca</td><td>ccgcccgtca</td><td>caccatggga</td><td>1380</td>

<td>gtcggaaatg</td><td>cccgaagtca</td><td>gtgacctaac</td><td>cgaaaggaag</td><td>gagctgccga</td><td>aggtggagcc</td><td>1440</td>

<td>ggtaactggg</td><td>gtgaagtcgt</td><td>aacaaggtag</td><td>CCGT</td><td></td><td></td><td>1474</td>

<210> 27 <211> 1484 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1484) <223> The sequence of the gene encoding the 16S rRNA 7 strain Clostridium <400> 27

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gagaatccag</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatctggag</td><td>gaaagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtga</td><td>gcaatctgcc</td><td>ttggagtggg</td><td>gaataacggt</td><td>tggaaacagc</td><td>cgctaatacc</td><td>180</td>

<td>gcatgatgcg</td><td>tctgggaggc</td><td>atctctctgg</td><td>acgccaaaga</td><td>tttatcgctc</td><td>tgagatgagc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagcttgtt</td><td>ggcggggtaa</td><td>aggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgcaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttctg</td><td>agggacgaag</td><td>aaagtgacgg</td><td>tacctcagga</td><td>480</td>

<td>ataagccacg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtaggtggc</td><td>aagcgttatc</td><td>540</td>

<td>cggatttatt</td><td>gggtgtaaag</td><td>ggcgtgtagg</td><td>cgggaaagca</td><td>agtcagatgt</td><td>gaaaactcag</td><td>600</td>

<td>ggctcaaccc</td><td>tgagcctgca</td><td>tttgaaactg</td><td>tttttcttga</td><td>gtgctggaga</td><td>ggcaatcgga</td><td>660</td>

<td>attccgtgtg</td><td>tagcggtgaa</td><td>atgcgtagat</td><td>atacggagga</td><td>caccagtggc</td><td>gagcggattg</td><td>720</td>

<td>ctggacagta</td><td>ctgacgctga</td><td>agcgcgaaag</td><td>cgtgggagca</td><td>aacagataga</td><td>tacctggtag</td><td>780</td>

<td>tcacgcgtaa</td><td>acgatggata</td><td>ctaggtgtgg</td><td>ggggactgac</td><td>cccctccgtg</td><td>ccgcagctaa</td><td>840</td>

<td>cgcaataagt</td><td>atcccacctg</td><td>gggagtacga</td><td>tcgcaaggtt</td><td>gaaactcaaa</td><td>ggaattgacg</td><td>900</td>

<td>ggggcccgca</td><td>caagcggtgg</td><td>agtatgtggt</td><td>ttaattcgaa</td><td>gcaacgcgaa</td><td>gaaccttacc</td><td>960</td>

<td>agggcttgac</td><td>atcctgctaa</td><td>cgaaccagag</td><td>atggattagg</td><td>tgcccttcgg</td><td>ggaaagcaga</td><td>1020</td>

<td>gacaggtggt</td><td>gcatggttgt</td><td>cgtcagctcg</td><td>tgtcgtgaga</td><td>tgttgggtta</td><td>agtcccgcaa</td><td>1080</td>

<td>cgagcgcaac</td><td>ccttattgtt</td><td>agttgctacg</td><td>caagagcact</td><td>ctagcgagac</td><td>tgccgttgac</td><td>1140</td>

<td>aaaacggagg</td><td>aaggtgggga</td><td>cgacgtcaaa</td><td>tcatcatgcc</td><td>ccttacgtcc</td><td>tgggccacac</td><td>1200</td>

<td>acgtactaca</td><td>atggcggcca</td><td>acaaagagag</td><td>gcaagaccgc</td><td>gaggtggagc</td><td>aaatctcaaa</td><td>1260</td>

<td>aagccgtccc</td><td>agttcggatc</td><td>gcaggctgca</td><td>acccgcctgc</td><td>gtgaagttgg</td><td>aatcgctagt</td><td>1320</td>

<td>aatcgcggat</td><td>cagcatgccg</td><td>cggtgaatac</td><td>gttcccgggc</td><td>cttgtacaca</td><td>ccgcccgtca</td><td>1380</td>

<td>caccatgaga</td><td>gtcgggaaca</td><td>cccgaagtcc</td><td>gtagcctaac</td><td>cgcaaggggg</td><td>gcgcggccga</td><td>1440</td>

<td>aggtgggttc</td><td>gataattggg</td><td>gtgaagtcgt</td><td>aacaaggtag</td><td>CCGT</td><td></td><td>1484</td>

<210> 28 <211> 1483 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1483) <223> The sequence of the gene encoding the 16S rRNA 8 of the Clostridium strain <400> 28

<td>agagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatgggaat</td><td>gcttagtggc</td><td>ggactggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>cttccagagg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcatgatgc</td><td>gttggagccg</td><td>catgactccg</td><td>acgtcaaaga</td><td>tttatcgctg</td><td>gaagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggtgaggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atggacgcaa</td><td>gtctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>ggggaagagc</td><td>agaagacggt</td><td>accccttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttgtcc</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gcgtgcagcc</td><td>ggagagacaa</td><td>gtcagatgtg</td><td>aaatccacgg</td><td>600</td>

<td>gctcaacccg</td><td>tgaactgcat</td><td>ttgaaactgt</td><td>ttcctttgag</td><td>tgtcggagag</td><td>gtaatcggga</td><td>660</td>

<td>ttccttgtgt</td><td>agcggtgaat</td><td>gcgtagatat</td><td>agagaccaca</td><td>gtgccgacgc</td><td>cgaatactga</td><td>720</td>

<td>cgatactgac</td><td>ggtgagcgcg</td><td>aaagcgtggg</td><td>gagcaaacag</td><td>gattagatac</td><td>cctggtagtc</td><td>780</td>

<td>cacgctgtaa</td><td>acgatcgata</td><td>ctaggtgtgc</td><td>ggggactgac</td><td>ccctgcgtgc</td><td>cggagttaac</td><td>840</td>

<td>acaataagta</td><td>tcgcacctgg</td><td>ggagtacgat</td><td>cgcaaggttg</td><td>aaactcaaag</td><td>gaattgacgg</td><td>900</td>

<td>gggcccgcac</td><td>aagcggtgga</td><td>ttatgtggtt</td><td>taattcgaag</td><td>caacgcgaag</td><td>aaccttacca</td><td>960</td>

<td>gggcttgaca</td><td>tcctgctaac</td><td>gaagtagaga</td><td>tacattaggt</td><td>gcccttcggg</td><td>gaaagcagag</td><td>1020</td>

<td>acaggtggtg</td><td>catggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>gttgggttaa</td><td>gtcccgcaac</td><td>1080</td>

<td>gagcgcaacc</td><td>cctattgtta</td><td>gttgctacgc</td><td>aagagcactc</td><td>tagcgagact</td><td>gccgttgaca</td><td>1140</td>

<td>aaacggagga</td><td>aggcggggac</td><td>gacgtcaaat</td><td>catcatgccc</td><td>cttatgtcct</td><td>gggctacaca</td><td>1200</td>

<td>cgtaatacaa</td><td>tggcggttaa</td><td>caaagggatg</td><td>caaagccgcg</td><td>aggcagagcg</td><td>aaccccaaaa</td><td>1260</td>

<td>agccgtccca</td><td>gttcggatcg</td><td>caggctgcaa</td><td>cccgcctgcg</td><td>tgaagtcgga</td><td>atcgctagta</td><td>1320</td>

<td>atcgcggatc</td><td>agcatgccgc</td><td>ggtgaatacg</td><td>ttcccgggcc</td><td>ttgtacacac</td><td>cgcccgtcac</td><td>1380</td>

<td>accatgagag</td><td>tcgggaacac</td><td>ccgaagtccg</td><td>tagcctaacc</td><td>gcaaggaggg</td><td>cgcggccgaa</td><td>1440</td>

<td>ggtgggttcg</td><td>ataattgggg</td><td>tgaagtcgta</td><td>acaaggtagc</td><td>cgt</td><td></td><td>1483</td>

<210> 29 <211> 1480 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1) ... (1480) <223> The sequence of the 16S rRNA gene for the 9 strain Clostridium <400> 29

<td>ggagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggctcatat</td><td>tgaaacctag</td><td>tgatgtatga</td><td>gttagtggcg</td><td>gacgggtgag</td><td>taacgcgtgg</td><td>120</td>

<td>agaacctgcc</td><td>gtatactggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>gcataagcgc</td><td>180</td>

<td>acagcttcgc</td><td>atgaagcagt</td><td>gtgaaaaact</td><td>ccggtggtat</td><td>acgatggatc</td><td>cgcgtctgat</td><td>240</td>

<td>tagctggttg</td><td>gcggggtaac</td><td>agcccaccaa</td><td>ggcgacgatc</td><td>agtagccggc</td><td>ctgagagggt</td><td>300</td>

<td>gaacggccac</td><td>attgggactg</td><td>agacacggcc</td><td>caaactccta</td><td>cgggaggcag</td><td>cagtggggaa</td><td>360</td>

<td>tattgcacaa</td><td>tgggggaaac</td><td>cctgatgcag</td><td>cgacgccgcg</td><td>tgagtgaaga</td><td>agtatttcgg</td><td>420</td>

<td>tatgtaaagc</td><td>tctatcagca</td><td>gggaagaaat</td><td>actgacctta</td><td>cggtcagcag</td><td>acggtacctg</td><td>480</td>

<td>actaagaagc</td><td>cccgggctaa</td><td>ctacgtgcca</td><td>gcagccgcgg</td><td>taatacgtag</td><td>gggcaagcgt</td><td>540</td>

<td>tatccggatt</td><td>tactgggtgt</td><td>aaagggggcg</td><td>cagacggcga</td><td>tgcaagccag</td><td>gagtgaaagc</td><td>600</td>

<td>cggggcccaa</td><td>ccccgggact</td><td>gctcttggac</td><td>tgcgtggctg</td><td>gagtgcagag</td><td>ggcagcgaat</td><td>660</td>

<td>tcctgtgtag</td><td>cgtgaatgcg</td><td>tagattcaga</td><td>ggacacgtgc</td><td>gagcgcctgc</td><td>tgactgcact</td><td>720</td>

<td>gacgtgagcc</td><td>cgaagcgtgg</td><td>ggagcaaaca</td><td>ggattagata</td><td>cctggtagtc</td><td>cacgccgtaa</td><td>780</td>

<td>acgatgatta</td><td>ctaggtgtcg</td><td>gggagcagag</td><td>actgcccggt</td><td>gccgcagcca</td><td>acgcattaag</td><td>840</td>

<td>taatccacct</td><td>ggggagtacg</td><td>ttcgcaagaa</td><td>tgaaactcaa</td><td>aggaattgac</td><td>ggggacccgc</td><td>900</td>

<td>acaagcggtg</td><td>gagcatgtgg</td><td>tttaattcga</td><td>agcaacgcga</td><td>agaaccttac</td><td>caggccttga</td><td>960</td>

<td>catccccctg</td><td>gatggcccgt</td><td>aacggggtca</td><td>gcctttcggg</td><td>gcaggggaga</td><td>caggtggtgc</td><td>1020</td>

<td>atggttgtcg</td><td>tcagctcgtg</td><td>tcgtgagatg</td><td>ttgggttaag</td><td>tcccgcaacg</td><td>agcgcaaccc</td><td>1080</td>

<td>ctgcccgcag</td><td>tagccagcat</td><td>tttagatggg</td><td>gactctgcgg</td><td>ggactgccgg</td><td>ggacaacccg</td><td>1140</td>

<td>gaggaaggcg</td><td>gggatgacgt</td><td>caaatcatca</td><td>tgccccttat</td><td>ggcctgggct</td><td>acacacgtgc</td><td>1200</td>

<td>tacaatggcg</td><td>ccgacagagg</td><td>gaggcgaagc</td><td>ggcgacgcgg</td><td>agcgaacccc</td><td>aaaaacggcg</td><td>1260</td>

<td>tcccagttcg</td><td>gattgtagtc</td><td>tgcaacccga</td><td>ctacatgaag</td><td>ccggaatcgc</td><td>tagtaatcgc</td><td>1320</td>

<td>ggatcagaat</td><td>gccgcggtga</td><td>atacgttccc</td><td>gggtcttgta</td><td>cacaccgccc</td><td>gtcacaccat</td><td>1380</td>

<td>gggagccggg</td><td>aatgcccgaa</td><td>gtctgtgacc</td><td>gaacccgtaa</td><td>ggggaggggc</td><td>agccgaaggc</td><td>1440</td>

<td>aggcccggtg</td><td>actggggtga</td><td>agtcgtaaca</td><td>aggtagccgt</td><td></td><td></td><td>1480</td>

<210> 30 <211> 1489 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1489) <223> The sequence of the gene encoding the 16S rRNA 10 strain Clostridium <400> 30

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgagc</td><td>60</td>

<td>gaagcacttt</td><td>tttagaactc</td><td>ttcggaggga</td><td>agagagggtg</td><td>acttagcggc</td><td>ggacgggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>ggcaacctgc</td><td>cttacacagg</td><td>gggataacaa</td><td>ttagaaatga</td><td>ttgctaatac</td><td>180</td>

<td>cgcataagac</td><td>cacggtactg</td><td>catggtacag</td><td>tggtaaaaac</td><td>tgaggtggtg</td><td>taagatgggc</td><td>240</td>

<td>ccgcgtctga</td><td>ttaggtagtt</td><td>ggtggggtag</td><td>aagcctacca</td><td>agccgacgat</td><td>cagtagccga</td><td>300</td>

<td>cctgagaggg</td><td>cgaccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccaaactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgcaca</td><td>atgggggaaa</td><td>ccctgatgca</td><td>gcgacgccgc</td><td>gtgagtgagg</td><td>420</td>

<td>aagtatttcg</td><td>gtatgtaaag</td><td>ctctatcagc</td><td>agggaagaaa</td><td>atgacggtac</td><td>ctgactaaga</td><td>480</td>

<td>agcccccggc</td><td>taactacgtg</td><td>ccagcagccg</td><td>cggtaatacg</td><td>tagggggcaa</td><td>gcgttatccg</td><td>540</td>

<td>gatttactgg</td><td>gtgtaaaggg</td><td>agcgtagacg</td><td>gacttgcaag</td><td>tctgatgtga</td><td>aaatccgggg</td><td>600</td>

<td>cccaacccgg</td><td>gactgcattg</td><td>aaactgtatt</td><td>ttttggaggg</td><td>gtccgaggag</td><td>gcaagtggaa</td><td>660</td>

<td>tcctgggtag</td><td>cggtgaaatg</td><td>gcgtagaatt</td><td>cagggaggaa</td><td>caccagtggc</td><td>ggaaggcgaa</td><td>720</td>

<td>ttactggacg</td><td>ataactgacg</td><td>gtgaggcgcg</td><td>aagcgtggga</td><td>gcaaacaaga</td><td>attagatacc</td><td>780</td>

<td>ctggtagtca</td><td>cgctgtaacg</td><td>atcgatacta</td><td>ggtgtgcggg</td><td>gactgacccc</td><td>tgcgtgccgg</td><td>840</td>

<td>agttaacaca</td><td>ataagtatcg</td><td>cactggggag</td><td>tacgatcgca</td><td>aggttgaaac</td><td>tcaaaggaat</td><td>900</td>

<td>tgacgggggc</td><td>ccgcacaagc</td><td>ggtggattat</td><td>gtggtttaat</td><td>tcgaagcaac</td><td>gcgaagaacc</td><td>960</td>

<td>ttaccagggc</td><td>ttgacatcct</td><td>gctaacgaag</td><td>tagagataca</td><td>ttaggtgccc</td><td>ttcggggaaa</td><td>1020</td>

<td>gcagagacag</td><td>gtggtgcatg</td><td>gttgtcgtca</td><td>gctcgtgtcg</td><td>tgagatgttg</td><td>ggttaagtcc</td><td>1080</td>

<td>cgcaacgagc</td><td>gcaaccccta</td><td>ttgttagttg</td><td>ctacgcaaga</td><td>gcactctagc</td><td>gagactgccg</td><td>1140</td>

<td>ttgacaaaac</td><td>ggaggaaggc</td><td>ggggacgacg</td><td>tcaaatcatc</td><td>atgcccctta</td><td>tgtcctgggc</td><td>1200</td>

<td>tacacacgta '</td><td>atacaatggc</td><td>ggttaacaaa</td><td>gggatgcaaa</td><td>gccgcgaggc</td><td>agagcgaacc</td><td>1260</td>

<td>ccaaaaagcc</td><td>gtcccagttc</td><td>ggatcgcagg</td><td>ctgcaacccg</td><td>cctgcgtgaa</td><td>gtcggaatcg</td><td>1320</td>

<td>ctagtaatcg</td><td>cggatcagca</td><td>tgccgcggtg</td><td>aatacgttcc</td><td>cgggccttgt</td><td>acacaccgcc</td><td>1380</td>

<td>cgtcacacca</td><td>tgagagtcgg</td><td>gaacacccga</td><td>agtccgtagc</td><td>ctaaccgcaa</td><td>ggagggcgcg</td><td>1440</td>

<td>gccgaaggtg</td><td>ggttcgataa</td><td>ttggggtgaa</td><td>gtcgtaacaa</td><td>ggtagccgt</td><td></td><td>1489</td>

<210> 31 <211> 1490 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1490) <223> The sequence of the gene encoding the 16S rRNA 11 strain Clostridium <400> 31

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gagaatccag</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatctggag</td><td>gaaagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtga</td><td>gcaatctgcc</td><td>ttggagtggg</td><td>gaataacggt</td><td>tggaaacagc</td><td>cgctaatacc</td><td>180</td>

<td>gcatgatgcg</td><td>tctgggaggc</td><td>atctctctgg</td><td>acgccaaaga</td><td>tttatcgctc</td><td>tgagatgagc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagcttgtt</td><td>ggcggggtaa</td><td>aggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgcaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttctg</td><td>agggacgaag</td><td>aaagtgacgg</td><td>tacctcagga</td><td>480</td>

<td>ataagccacg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtaggtggc</td><td>aagcgttatc</td><td>540</td>

<td>cggatttatt</td><td>gggtgtaaag</td><td>ggcgtgtagg</td><td>cgggaaagca</td><td>agtcagatgt</td><td>gaaaactcag</td><td>600</td>

<td>ggctcaaccc</td><td>tgagcctgca</td><td>tttgaaactg</td><td>tttttcttga</td><td>gtgctggaga</td><td>ggcaatcgga</td><td>660</td>

<td>attccgtgtt</td><td>gtagcggtga</td><td>aatgcgtaga</td><td>ttataccgga</td><td>ggaaccacca</td><td>gtggcggaag</td><td>720</td>

<td>gcggattgct</td><td>ggaacagtaa</td><td>ctgacgctga</td><td>ggcgccgaaa</td><td>gcgtggggag</td><td>caaacaggat</td><td>780</td>

<td>agataccctg</td><td>gtagtccacg</td><td>ccgtaaacga</td><td>tggatactaa</td><td>gtgtggggga</td><td>ctgacccctt</td><td>840</td>

<td>cgtgcccagc</td><td>taagcaataa</td><td>gtttcccacc</td><td>tggggagtac</td><td>gatcgcaggt</td><td>gaaactcaaa</td><td>900</td>

<td>ggaattgacg</td><td>ggggcccgcc</td><td>caagcgggtg</td><td>gagtaggggt</td><td>taattggagc</td><td>aacgggaaga</td><td>960</td>

<td>accttaccag</td><td>ggcttgacat</td><td>cctgtaacga</td><td>accagaagag</td><td>ggattaggtg</td><td>ccttcgggga</td><td>1020</td>

<td>aagcagagac</td><td>aggtggtgca</td><td>tggttgtcgt</td><td>cagctcgtgt</td><td>cgtgagatgt</td><td>gggtaaagtc</td><td>1080</td>

<td>ccgcaacgag</td><td>cgcaaccctt</td><td>attgttagtt</td><td>gctacgcaag</td><td>agcactctag</td><td>cgagactgcc</td><td>1140</td>

<td>gttgacaaaa</td><td>cggaggaagg</td><td>tggggacgac</td><td>gtcaaatcat</td><td>catgcccctt</td><td>acgtcctggg</td><td>1200</td>

<td>ccacacacgt</td><td>actacaatgg</td><td>cggccaacaa</td><td>agagaggcaa</td><td>gaccgcgagg</td><td>tggagaaaat</td><td>1260</td>

<td>ctcaaaaagc</td><td>cgtcccagtt</td><td>cggatcgcag</td><td>gctgcaaccc</td><td>gcctgcgtga</td><td>agttggaatc</td><td>1320</td>

<td>gctagtaatc</td><td>gcggatcagc</td><td>atgccgcggt</td><td>gaatacgttc</td><td>ccgggccttg</td><td>tacacaccgc</td><td>1380</td>

<td>ccgtcacacc</td><td>atgagagtcg</td><td>ggaacacccg</td><td>aagtccgtag</td><td>cctaaccgca</td><td>aggggggcgc</td><td>1440</td>

<td>ggccgaaggt</td><td>gggttcgata</td><td>attggggtga</td><td>agtcgtaaca</td><td>aggtagccgt</td><td></td><td>1490</td>

<210> 32 <211> 1489 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1) ... (1489) <223> The sequence of the 16S rRNA gene for the 12 strain of Clostridium <400> 32

<td>agagtttgat</td><td>catggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggctcatat</td><td>tgaaacctag</td><td>tgatgtatga</td><td>gttagtggcg</td><td>gacgggtgag</td><td>taacgcgtgg</td><td>120</td>

<td>agaacctgcc</td><td>gtatactggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>gcataagcgc</td><td>180</td>

<td>acagcttcgc</td><td>atgaagcagt</td><td>gtgaaaaact</td><td>ccggtggtat</td><td>acgatggatc</td><td>cgcgtctgat</td><td>240</td>

<td>tagctggttg</td><td>gcggggtaac</td><td>agcccaccaa</td><td>ggcgacgatc</td><td>agtagccggc</td><td>ctgagagggt</td><td>300</td>

<td>gaacggccac</td><td>attgggactg</td><td>agacacggcc</td><td>caaactccta</td><td>cgggaggcag</td><td>cagtggggaa</td><td>360</td>

<td>tattgcacaa</td><td>tgggggaaac</td><td>cctgatgcag</td><td>cgacgccgcg</td><td>tgagtgaaga</td><td>agtatttcgg</td><td>420</td>

<td>tatgtaaagc</td><td>tctatcagca</td><td>gggaagaaat</td><td>actgacctta</td><td>cggtcagcag</td><td>acggtacctg</td><td>480</td>

<td>actaagaagc</td><td>cccggctaac</td><td>tacgtgccag</td><td>cagccgcggt</td><td>aatacgtagg</td><td>gggcaagcgt</td><td>540</td>

<td>tatccggatt</td><td>tactgggtgt</td><td>aaagggagcg</td><td>tagacggcag</td><td>cgcaagtctg</td><td>aagtgaaatc</td><td>600</td>

<td>ccatggctta</td><td>accatggaac</td><td>tgctttggaa</td><td>actgtgcagc</td><td>tggagtgcag</td><td>gagaggtaag</td><td>660</td>

<td>cggaattcct</td><td>agtgtagcgg</td><td>tgaatgcgta</td><td>gatattagag</td><td>gacaccagtg</td><td>gcgatgcggc</td><td>720</td>

<td>ttactggact</td><td>gtactgacgt</td><td>tgagctcgaa</td><td>agcgtgggga</td><td>gcaccagaat</td><td>tagaatactg</td><td>780</td>

<td>tagtcacgcc</td><td>gtaaccgatg</td><td>atactaggtg</td><td>tgggggacca</td><td>aggtctcgtg</td><td>ccggcggcaa</td><td>840</td>

<td>acgcattaag</td><td>taatccacct</td><td>ggggagtacg</td><td>ttcgcaagaa</td><td>tgaaactcaa</td><td>aggaattgac</td><td>900</td>

<td>ggggacccgc</td><td>acaagcggtg</td><td>gagcatgtgg</td><td>tttaattcga</td><td>agcaacgcga</td><td>agaaccttac</td><td>960</td>

<td>ctggtcttga</td><td>catcccgatg</td><td>acgagtgagc</td><td>aaagtcactt</td><td>tcccttcggg</td><td>gcattggaga</td><td>1020</td>

<td>caggtggtgc</td><td>atggttgtcg</td><td>tcagctcgtg</td><td>tcgtgagatg</td><td>ttgggttaag</td><td>tcccgcaacg</td><td>1080</td>

<td>agcgcaaccc</td><td>ctatttccag</td><td>tagccagcag</td><td>gtagagctgg</td><td>gcactctgga</td><td>gagactgccc</td><td>1140</td>

<td>gggataaccg</td><td>ggaggaaggc</td><td>ggggatgacg</td><td>tcaaatcatc</td><td>atgcccctta</td><td>tgatcagggc</td><td>1200</td>

<td>tacacacgtg</td><td>ctacaatggc</td><td>gtaaacaaag</td><td>ggaagcgaga</td><td>cggtgacgtt</td><td>aagcaaatcc</td><td>1260</td>

<td>caaaaataac</td><td>gtcccagttc</td><td>ggattgtagt</td><td>ctgcaactcg</td><td>actacatgaa</td><td>gctggaatcg</td><td>1320</td>

ctagtaatcg cgaatcagaa tgtcgcggtg aatacgttcc cgggtcttgt acacaccgcc 1380 cgtcacacca tgggagtcgg aaatgcccga agtcagtgac ctaaccgaaa ggaaggagct 1440 gccgaaggtg gagccggtaa ctggggtgaa gtcgtaacaa ggtagccgt 1489 <210> 33 <211> 1456 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1) ... (1456) <223> The sequence of the gene encoding 16S rRNA 13 of the Clostridium strain

-63 <400> 33

<td>agagtttgat</td><td>catggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gaagcacttg</td><td>agaacgattc</td><td>ttcggatgag</td><td>gacttttgtg</td><td>actgagtggc</td><td>ggacgggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>ggtaacctgc</td><td>cctatacagg</td><td>gggataacag</td><td>ttagaaatga</td><td>ctgctaatac</td><td>180</td>

<td>cgcataagcg</td><td>cactaaaacc</td><td>gcatggttcg</td><td>gtgtgaaaaa</td><td>ctgaggtggt</td><td>ataggatgga</td><td>240</td>

<td>cccgcgtctg</td><td>attagcttgt</td><td>tggtggggta</td><td>acggctcacc</td><td>aaggcgacga</td><td>tcagtagccg</td><td>300</td>

<td>gcctgagagg</td><td>gcgaccggcc</td><td>acattgggac</td><td>tgagacacgg</td><td>cccaaactcc</td><td>tacgggaggc</td><td>360</td>

<td>agcagtgggg</td><td>gatattgcac</td><td>aatgggggga</td><td>accctgatgc</td><td>agcgacgccg</td><td>cgtgggtgaa</td><td>420</td>

<td>gaagcgcctc</td><td>ggcgcgtaaa</td><td>gccctgtcag</td><td>cagggaagaa</td><td>aatgacggta</td><td>cctgaagaag</td><td>480</td>

<td>aagccccggc</td><td>taactacgtg</td><td>ccagcagccg</td><td>cggtaatacg</td><td>taggggcaag</td><td>cgttattccg</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gggcgcagac</td><td>ggcgatgcaa</td><td>gccaggagtg</td><td>aagcccgggg</td><td>600</td>

<td>cccacccggg</td><td>actgctcttg</td><td>gactgcgtgc</td><td>tggagtgcag</td><td>aaggggcagc</td><td>gatcctgtgt</td><td>660</td>

<td>accgtgaatt</td><td>gcgtagatat</td><td>cagagacacg</td><td>ttgcgagcgc</td><td>tgctgactgc</td><td>actgacgtga</td><td>720</td>

<td>gcgaagctgg</td><td>agcacagata</td><td>gatactgtag</td><td>tcagcgtaac</td><td>gatgatacta</td><td>gtgtcgggag</td><td>780</td>

<td>cagagactgc</td><td>ccgttgcggc</td><td>agcccaacgc</td><td>attagtattc</td><td>cacttgggga</td><td>gtacgtttcg</td><td>840</td>

<td>cagaatgaac</td><td>ttcaaggaaa</td><td>tgacggggac</td><td>ccgcacaagg</td><td>cggtggagca</td><td>tgtggtttaa</td><td>900</td>

<td>ttcgaagcaa</td><td>cgcgaagaac</td><td>cttaccaggc</td><td>cttgacatcc</td><td>cccctggatg</td><td>gcccgtaacg</td><td>960</td>

<td>gggtcagcct</td><td>ttcggggcag</td><td>gggagacagg</td><td>tggtgcatgg</td><td>ttgtcgtcag</td><td>ctcgtgtcgt</td><td>1020</td>

<td>gagatgttgg</td><td>gttaagtccc</td><td>gcaacgagcg</td><td>caacccctgc</td><td>ccgcagtagc</td><td>cagcatttta</td><td>1080</td>

<td>gatggggact</td><td>ctgcggggac</td><td>tgccggggac</td><td>aacccggagg</td><td>aaggcgggga</td><td>tgacgtcaaa</td><td>1140</td>

<td>tcatcatgcc</td><td>ccttatggcc</td><td>tgggctacac</td><td>acgtgctaca</td><td>atggcgccga</td><td>cagagggagg</td><td>1200</td>

<td>cgaagcggcg</td><td>acgcggagcg</td><td>aaccccaaaa</td><td>acggcgtccc</td><td>agttcggatt</td><td>gtagtctgca</td><td>1260</td>

<td>acccgactac</td><td>atgaagccgg</td><td>aatcgctagt</td><td>aatcgcggat</td><td>cagaatgccg</td><td>cggtgaatac</td><td>1320</td>

<td>gttcccgggt</td><td>cttgtacaca</td><td>ccgcccgtca</td><td>caccatggga</td><td>gccgggaatg</td><td>cccgaagtct</td><td>1380</td>

<td>gtgaccgaac</td><td>ccgtaagggg</td><td>aggggcagcc</td><td>gaaggcaggc</td><td>tcggtgactg</td><td>gggtgaagtc</td><td>1440</td>

<td>gtaacaaggt</td><td>agccgt</td><td></td><td>.... _______</td><td></td><td></td><td>1456</td>

<210> 34 <211> 1475 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1475) <223> The sequence of the 16S rRNA gene for the 14 strain Clostridium

-64 <400> 34

<td>agagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatgggaat</td><td>gcttagtggc</td><td>ggactggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>cttccagagg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcatgatgc</td><td>gttggagccg</td><td>catgactccg</td><td>acgtcaaaga</td><td>tttatcgctg</td><td>gaagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggtgaggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atggacgcaa</td><td>gtctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>ggggaagagc</td><td>agaagacggt</td><td>accccttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttgtcc</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gcgtgcagcc</td><td>ggagagacaa</td><td>gtcagatgtg</td><td>aaatccacgg</td><td>600</td>

<td>gctcaacccg</td><td>tgaactgcat</td><td>ttgaaactgt</td><td>ttcccttgag</td><td>tgtcggagag</td><td>gtaatcggaa</td><td>660</td>

<td>tttccttgtg</td><td>tagcggtgaa</td><td>tgcgtagata</td><td>taaggaagga</td><td>cacagtggcg</td><td>agcggattac</td><td>720</td>

<td>tggacgatac</td><td>tgacgtgagc</td><td>gcgaaagcgt</td><td>gggggagcaa</td><td>cagaaattag</td><td>atactgtagt</td><td>780</td>

<td>gcagctgtaa</td><td>cgatcgatac</td><td>tagttgcggg</td><td>actgacccct</td><td>tgcgtgcgag</td><td>ttacacaata</td><td>840</td>

<td>agtatcgcac</td><td>ctgggagtac</td><td>gatcgcaagg</td><td>ttggaactca</td><td>aaggaattga</td><td>cggggcccgc</td><td>900</td>

<td>acaagcgttg</td><td>gattatgtgg</td><td>tttaattcga</td><td>agcaacgcga</td><td>agaaccttac</td><td>cagggcttga</td><td>960</td>

<td>catcctgcta</td><td>acgaagtaga</td><td>gatacattag</td><td>gtgcccttcg</td><td>gggaaagtag</td><td>agacaggtgg</td><td>1020</td>

<td>tgcatggttg</td><td>tcgtcagctc</td><td>gtgtcgtgag</td><td>atgttgggtt</td><td>aagtcccgca</td><td>acgagcgcaa</td><td>1080</td>

<td>cccctattgt</td><td>tagttgctac</td><td>gcaagagcac</td><td>tctagcgaga</td><td>ctgccgttga</td><td>caaaacggag</td><td>1140</td>

<td>gaaggcgggg</td><td>acgacgtcaa</td><td>atcatcatgc</td><td>cccttatgtc</td><td>ctgggctaca</td><td>cacgtaatac</td><td>1200</td>

<td>aatggcggtt</td><td>aacaaaggga</td><td>tgcaaagccg</td><td>cgaggcagag</td><td>cgaaccccaa</td><td>aaagccgtcc</td><td>1260</td>

<td>cagttcggat</td><td>cgcaggctgc</td><td>aacccgcctg</td><td>cgtgaagtcg</td><td>gaatcgctag</td><td>taatcgcgga</td><td>1320</td>

<td>tcagcatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>ccttgtacac</td><td>accgcccgtc</td><td>acaccatgag</td><td>1380</td>

<td>agtcgggaac</td><td>acccgaagtc</td><td>cgtagcctaa</td><td>ccgcaaggag</td><td>ggcgcggccg</td><td>aaggtgggtt</td><td>1440</td>

<td>cgataattgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td></td><td></td><td>1475</td>

<210> 35 <211> 1480 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1) ... (1480) <223> The sequence of the gene encoding 16S rRNA 15 of the Clostridium strain

-65 <400> 35

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggctcatat</td><td>tgaaacctag</td><td>tgatgtatga</td><td>gttagtggcg</td><td>gacgggtgag</td><td>taacgcgtgg</td><td>120</td>

<td>agaacctgcc</td><td>gtatactggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>gcataagcgc</td><td>180</td>

<td>acagcttcgc</td><td>atgaagcagt</td><td>gtgaaaaact</td><td>ccggtggtat</td><td>acgatggatc</td><td>cgcgtctgat</td><td>240</td>

<td>tagctggttg</td><td>gcggggtaac</td><td>agcccaccaa</td><td>ggcgacgatc</td><td>agtagccggc</td><td>ctgagagggt</td><td>300</td>

<td>gaacggccac</td><td>attgggactg</td><td>agacacggcc</td><td>caaactccta</td><td>cgggaggcag</td><td>cagtggggaa</td><td>360</td>

<td>tattgcacaa</td><td>tgggggaaac</td><td>cctgatgcag</td><td>cgacgccgcg</td><td>tgagtgaaga</td><td>agtatttcgg</td><td>420</td>

<td>tatgtaaagc</td><td>tctatcagca</td><td>gggaagaaat</td><td>actgacctta</td><td>cggtcagcag</td><td>acggtacctg</td><td>480</td>

<td>actaagaagc</td><td>cccggctaac</td><td>tacgtgccag</td><td>cagccgcggt</td><td>aatacgtagg</td><td>ggcaagcgtt</td><td>540</td>

<td>atccggattt</td><td>actgggtgta</td><td>aagggagcgt</td><td>agacggcagc</td><td>gcaagtctga</td><td>agtgaaatcc</td><td>600</td>

<td>catggcttaa</td><td>cccatggaac</td><td>tgctttggaa</td><td>actgtgcagc</td><td>tggagtgcag</td><td>gagaggtaag</td><td>660</td>

<td>cggaattcct</td><td>agtgtagcgt</td><td>gaaatgcgta</td><td>gattattagg</td><td>aggacaacag</td><td>tgcgagcgct</td><td>720</td>

<td>actgacgtga</td><td>ggctcgaagc</td><td>gtgggagcaa</td><td>acaggattag</td><td>atacctggta</td><td>gtcacgcgta</td><td>780</td>

<td>aacgatgatt</td><td>actagggtgt</td><td>tgggggacca</td><td>aggtcttcgg</td><td>tgccggcgca</td><td>aacgcattaa</td><td>840</td>

<td>gtaatccacc</td><td>tggggagtac</td><td>gttcgcaaga</td><td>atgaaactca</td><td>aaggaattga</td><td>cggggacccg</td><td>900</td>

<td>cacaagcggt</td><td>ggagcatgtg</td><td>gtttaattcg</td><td>aagcaacgcg</td><td>aagaacctta</td><td>cctggtcttg</td><td>960</td>

<td>acatcccgat</td><td>gacgagtgag</td><td>caaagtcact</td><td>ttcccttcgg</td><td>ggcattggag</td><td>acaggtggtg</td><td>1020</td>

<td>catggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>gttgggttaa</td><td>gtcccgcaac</td><td>gagcgcaacc</td><td>1080</td>

<td>cctatttcca</td><td>gtagccagca</td><td>ggtagagctg</td><td>ggcactctgg</td><td>agagactgcc</td><td>cgggataacc</td><td>1140</td>

<td>gggaggaagg</td><td>cggggatgac</td><td>gtcaaatcat</td><td>catgcccctt</td><td>atgatcaggg</td><td>ctacacacgt</td><td>1200</td>

<td>gctacaatgg</td><td>cgtaaacaaa</td><td>gggaagcgag</td><td>acggtgacgt</td><td>taagcaaatc</td><td>ccaaaaataa</td><td>1260</td>

<td>cgtcccagtt</td><td>cggattgtag</td><td>tctgcaactc</td><td>gactacatga</td><td>agctggaatc</td><td>gctagtaatc</td><td>1320</td>

<td>gcgaatcaga</td><td>atgtcgcggt</td><td>gaatacgttc</td><td>ccgggtcttg</td><td>tacacaccgc</td><td>ccgtcacacc</td><td>1380</td>

<td>atgggagtcg</td><td>gaaatgcccg</td><td>aagtcagtga</td><td>cctaaccgaa</td><td>aggaaggagc</td><td>tgccgaaggt</td><td>1440</td>

<td>ggagccggta</td><td>actggggtga</td><td>agtcgtaaca</td><td>aggtagccgt</td><td></td><td></td><td>1480</td>

<210> 36 <211> 1486 <212> DNA <213> Clostridium papyrosolvens <220>

<221> rRNA <222> (1) ... (1486) <223> The sequence of the 16S rRNA gene 16 strains of Clostridium

-66 <400> 36

<td>agagtttgat</td><td>cctggctcag</td><td>gataaacgct</td><td>ggcggcgcac</td><td>ataagacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggacttaact</td><td>cattctttta</td><td>gattgagagc</td><td>ggttagtggc</td><td>ggactggtga</td><td>gtaacacgta</td><td>120</td>

<td>agcaacctgc</td><td>ctatcagagg</td><td>ggaataacag</td><td>tgagaaatca</td><td>ttgctaatac</td><td>cgcatatgct</td><td>180</td>

<td>cacagtatca</td><td>catgatacag</td><td>tgaggaaagg</td><td>agcaatccgc</td><td>tgatagatgg</td><td>gcttgcgcct</td><td>240</td>

<td>gattagttag</td><td>ttggtggggt</td><td>aacggcctac</td><td>caagacgacg</td><td>atcagtagcc</td><td>ggactgagag</td><td>300</td>

<td>gttgaacggc</td><td>cacattggga</td><td>ctgagatacg</td><td>gcccagactc</td><td>ctacgggagg</td><td>cagcagtcgg</td><td>360</td>

<td>gaatattgcg</td><td>caatggagga</td><td>aactctgacg</td><td>cagtgacgcc</td><td>gcgtatagga</td><td>agaaggtttt</td><td>420</td>

<td>cggattgtaa</td><td>actattgtcg</td><td>ttagggaaga</td><td>taaaagactg</td><td>tacctaagga</td><td>ggaagccccg</td><td>480</td>

<td>gctaactatg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>catagggggc</td><td>aagcgttatc</td><td>cggaattatt</td><td>540</td>

<td>gggtgtaaag</td><td>ggtgcgtaga</td><td>cggaagaaca</td><td>agttggttgt</td><td>gaaatccctc</td><td>ggctcaactg</td><td>600</td>

<td>aggaactgca</td><td>accaaaacta</td><td>ttctccttga</td><td>gtgtcggaga</td><td>ggaaagtgga</td><td>attcctagtg</td><td>660</td>

<td>tagcggtgaa</td><td>atgcgtagat</td><td>attaggagga</td><td>acaccagtgg</td><td>cgaaggcgac</td><td>tttctggacg</td><td>720</td>

<td>ataactgacg</td><td>ttgaggcacg</td><td>aaagtgtggg</td><td>gagcaaacag</td><td>gattagatac</td><td>cctggtagtc</td><td>780</td>

<td>cacactgtaa</td><td>acgatggata</td><td>ctaggtgtag</td><td>ggtgtattaa</td><td>gcactctgtg</td><td>ccgccg.ctaa</td><td>840</td>

<td>cgcattaagt</td><td>atcccacctg</td><td>gggagtacga</td><td>ccgcaaggtt</td><td>gaaactcaaa</td><td>ggaattgacg</td><td>900</td>

<td>ggggcccgca</td><td>caagcagtgg</td><td>agtatgtggt</td><td>ttaattcgaa</td><td>gcaacgcgaa</td><td>gaaccttacc</td><td>960</td>

<td>agggcttgac</td><td>atataccgga</td><td>atatactaga</td><td>gatagtatag</td><td>tccttcggga</td><td>ctggtataca</td><td>1020</td>

<td>ggtggtgcat</td><td>ggttgtcgtc</td><td>agctcgtgtc</td><td>gtgagatgtt</td><td>gggttaagtc</td><td>ccgcaacgag</td><td>1080</td>

<td>cgcaacccct</td><td>atcgttagtt</td><td>gctagcaggt</td><td>aatgctgaga</td><td>actctagcga</td><td>gactgccggt</td><td>1140</td>

<td>gataaatcgg</td><td>aggaaggtgg</td><td>ggatgacgtc</td><td>aaatcatcat</td><td>gccctttatg</td><td>tcctgggcta</td><td>1200</td>

<td>cacacgtact</td><td>acaatggccg</td><td>taacagaggg</td><td>aagcaatata</td><td>gtgatatgga</td><td>gcaaaaccct</td><td>1260</td>

<td>aaaagcggtc</td><td>tcagttcgga</td><td>ttgaaggctg</td><td>aaattcgcct</td><td>tcatgaagcc</td><td>ggaattgcta</td><td>1320</td>

<td>gtaatggcag</td><td>gtcagcatac</td><td>tgccgtgaat</td><td>acgttcccgg</td><td>gccttgtaca</td><td>caccgcccgt</td><td>1380</td>

<td>cacaccatga</td><td>gagttggaaa</td><td>tacccgaagc</td><td>ctgtgagcta</td><td>actgtaaaga</td><td>ggcagcagtc</td><td>1440</td>

<td>gaaggtagag</td><td>ccaatgattg</td><td>gggtgaagtc</td><td>gtaacaaggt</td><td>agccgt</td><td></td><td>1486</td>

<210> 37 <211> 1493 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1493) <223> The sequence of the 16S rRNA gene for the 17 strain of Clostridium <400> 37

-67agagtttgat catggctcag gatgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 gagaaccaac ggattgagga ttcgtccaaa tgaagttggg gaaagtggcg gacgggtgag 120 taacgcgtga gcaatctgcc ttggagtggg gaataacggt tggaaacagc cgctaatacc 180

<td>gcatgatgcg</td><td>tctgggaggc</td><td>atctctctgg</td><td>acgccaaaga</td><td>tttatcgctc</td><td>tgagatgagc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggcggggcaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgcaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>ggggacgaac</td><td>aaatgacggt</td><td>accccttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttatcc</td><td>540</td>

<td>ggatttattg</td><td>ggtgtaaagg</td><td>gcgtgtaggc</td><td>gggaatgcaa</td><td>gtcagatgtg</td><td>aaaactatgg</td><td>600</td>

<td>gctcaaccca</td><td>tagcctgcat</td><td>ttgaaactgt</td><td>atttcttgag</td><td>tgctggagag</td><td>gcaatcggaa</td><td>660</td>

<td>ttccgtgtgt</td><td>agcggtgaaa</td><td>tgcgtagata</td><td>tacggaggaa</td><td>caccagtggc</td><td>gaagcggatt</td><td>720</td>

<td>gctggacagt</td><td>aactgacgct</td><td>gaggcgcgaa</td><td>agcgtgggga</td><td>gcaaacaggg</td><td>attagatacc</td><td>780</td>

<td>ctggtagtca</td><td>cgccgtaaac</td><td>gatggatact</td><td>aggtgtgggg</td><td>ggactgaccc</td><td>cctccgtgcc</td><td>840</td>

<td>gcagctaacg</td><td>caataagtat</td><td>cccacctggg</td><td>gagtacgatc</td><td>gcaagggttg</td><td>aaactcaaag</td><td>900</td>

<td>gaattgacgg</td><td>gggcccgcac</td><td>aagcggtgga</td><td>gtatgtggtt</td><td>taattcgaag</td><td>caacgcgaag</td><td>960</td>

<td>aaccttacca</td><td>gggcttgaca</td><td>tcctgctaac</td><td>gaaccagaga</td><td>tggatcaggt</td><td>gcccttcggg</td><td>1020</td>

<td>gaaagcagag</td><td>acaggtggtg</td><td>catggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>gttgggttaa</td><td>1080</td>

<td>gtcccgcaac</td><td>gagcgcaacc</td><td>cctattgtta</td><td>gttgctacgc</td><td>aagagcactc</td><td>tagcgagact</td><td>1140</td>

<td>gccgttgaca</td><td>aaacggagga</td><td>aggtggggac</td><td>gacgtcaaat</td><td>catcatgccc</td><td>cttacgtcct</td><td>1200</td>

<td>gggccacaca</td><td>cgtactacaa</td><td>tggcggccaa</td><td>caaagagagg</td><td>caagaccgcg</td><td>aggtggagca</td><td>1260</td>

<td>aatctcaaaa</td><td>agccgtccca</td><td>gttcggatcg</td><td>caggctgcaa</td><td>cccgcctgcg</td><td>tgaagttgga</td><td>1320</td>

<td>atcgctagta</td><td>atcgcggatc</td><td>agcatgccgc</td><td>ggtgaatacg</td><td>ttcccgggcc</td><td>ttgtacacac</td><td>1380</td>

<td>cgcccgtcac</td><td>accatgagag</td><td>tcgggaacac</td><td>ccgaagtccg</td><td>tagcctgacc</td><td>gcaagggggg</td><td>1440</td>

<td>cgcggccgaa</td><td>ggtgggttcg</td><td>ataattgggg</td><td>tgaagtagta</td><td>acaaggtagc</td><td>cgt</td><td>1493</td>

<210> 38 <211> 1493 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1) ... (1493) <223> The sequence of the gene encoding the 16S rRNA of the 18 strain Clostridium <400> 38

<td>agagtttgat</td><td>cctggctcag gacgaacgct ggcggcgcgc ctaacacatg caagtcgaac</td><td>60</td>

<td>ggagcttata</td><td>tttcagaagt tttcggatgg acgagagata agcttagtgg cggacgggtg</td><td>120</td>

<td>agtaacacgt</td><td>gagcaacctg cctttcagag ggggataaca gttggaaacg actgctaata</td><td>180</td>

<td>ccgcataacg</td><td>ctgcgatggg gcatcccgat gcagccaaag gagcaatccg ctgaaagatg</td><td>240</td>

<td>ggctcgcggc</td><td>cgattagcta gttggtgggg caacggccca ccaaggcgac gatcggtagc</td><td>300</td>

<td>cggactgaga</td><td>ggttgatcgg</td><td>ccacattggg</td><td>actgagacac</td><td>ggcccagact</td><td>cctacgggag</td><td>360</td>

<td>gcagcagtgg</td><td>gggatattgc</td><td>acaatggagg</td><td>aaactctgat</td><td>gcagcgacgc</td><td>cgcgtgaggg</td><td>420</td>

<td>aagacggtct</td><td>tcggattgta</td><td>aacctctgtc</td><td>tttggggaag</td><td>aaaatgacgg</td><td>tacccaaaga</td><td>480</td>

<td>ggaagctccg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtacggagc</td><td>gagcgttgtc</td><td>540</td>

<td>cggaattact</td><td>gggtgtaaag</td><td>ggagcgtacg</td><td>cgggcgagaa</td><td>agttgaatgt</td><td>taaatctacc</td><td>600</td>

<td>ggcttaactg</td><td>gtagctgcgt</td><td>tcaaaacttc</td><td>ttgtcttgag</td><td>tgaagtagag</td><td>gcaggcggaa</td><td>660</td>

<td>ttcctagtgt</td><td>agcggtgaaa</td><td>tgcgtagata</td><td>taggaggaca</td><td>ccagtgggcg</td><td>aagccgcctg</td><td>720</td>

<td>ctgggcttta</td><td>actgacgctg</td><td>aggctcgaaa</td><td>gcgtggggag</td><td>caaaccagga</td><td>ttagataccc</td><td>780</td>

<td>tggtagtcaa</td><td>cgctgtaaac</td><td>gatgattact</td><td>aggtgtgggg</td><td>gggactgacc</td><td>ccctccgtgc</td><td>840</td>

<td>cgcagttaac</td><td>acaataagta</td><td>tccacctggg</td><td>gagtacggcc</td><td>gcaaagtttg</td><td>aaaactcaaa</td><td>900</td>

<td>aggaatgacg</td><td>ggggcccgca</td><td>caaagcagtg</td><td>gagtatgtgg</td><td>tttaatttcg</td><td>aagcaacgcg</td><td>960</td>

<td>aagaacctta</td><td>ccaggtcttg</td><td>acatcgtgcg</td><td>catagcctag</td><td>agataggtga</td><td>agcccttcgg</td><td>1020</td>

<td>ggcgcacaga</td><td>caggtggtgc</td><td>atggttgtcg</td><td>tcagctcgtg</td><td>tcgtgagatg</td><td>ttgggttaag</td><td>1080</td>

<td>tcccgcaacg</td><td>agcgcaaccc</td><td>ttattattag</td><td>ttgctacgca</td><td>agagcactct</td><td>aatgagactg</td><td>1140</td>

<td>ccgttgacaa</td><td>aacggaggaa</td><td>ggtggggatg</td><td>acgtcaaatc</td><td>atcatgcccc</td><td>ttatgacctg</td><td>1200</td>

<td>ggctacacac</td><td>gtactacaat</td><td>ggcactgaaa</td><td>cagagggaag</td><td>cgacatcgcg</td><td>aggtgaagcg</td><td>1260</td>

<td>aatcccaaaa</td><td>aagtgtccca</td><td>gttcggattg</td><td>caggctgcaa</td><td>ctcgcctgca</td><td>tgaagtcgga</td><td>1320</td>

<td>attgctagta</td><td>atcgcggatc</td><td>agcatgccgc</td><td>ggtgaatacg</td><td>ttcccgggcc</td><td>ttgtacacac</td><td>1380</td>

<td>cgcccgtcac</td><td>accatgggag</td><td>tcggtaacac</td><td>ccgaagccag</td><td>tagcctaacc</td><td>gcaaggaggg</td><td>1440</td>

<td>cgctgtcgaa</td><td>ggtgggattg</td><td>atgactgggg</td><td>tgaagtcgta</td><td>acaaggtagc</td><td>cgt</td><td>1493</td>

<210> 39 <211> 1483 <212> DNA <213> Clostridium leptum <220>

<221> właściwość różna <222> (1)..(1483) <223> Sekwencja genu kodującego 16S rRNA 19 szczepu Clostridium <400> 39

<td>agagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcacgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagtgaaga</td><td>tgcttgcatc</td><td>tgaacttagt</td><td>ggcggacggg</td><td>tgagtaacac</td><td>gtgagcaacc</td><td>120</td>

<td>tgcctttcag</td><td>agggggataa</td><td>cgtttggaaa</td><td>cgaacgctaa</td><td>taccgcataa</td><td>aatatcggag</td><td>180</td>

<td>tcgcatggca</td><td>ctgatatcaa</td><td>aggagcaatc</td><td>cgctgaaaga</td><td>tgggctcgcg</td><td>tccgattagg</td><td>240</td>

<td>cagttggcgg</td><td>ggtaacggcc</td><td>caccaaaccg</td><td>acaatcggta</td><td>gccggactga</td><td>gaggttgaac</td><td>300</td>

<td>ggccacattg</td><td>ggactgagac</td><td>acggcccaga</td><td>ctcctacggg</td><td>aggcagcagt</td><td>gggggatatt</td><td>360</td>

<td>gcacaatggg</td><td>ggaaaccctg</td><td>atgcagcgat</td><td>gccgcgtgaa</td><td>tgaagacggc</td><td>cttcgggttg</td><td>420</td>

<td>taaagttctg</td><td>tcgcagggga</td><td>cgaaaatgac</td><td>ggtaccctgc</td><td>aagaaagctc</td><td>cggctaacta</td><td>480</td>

<td>cgtgccagca</td><td>gccgcggtaa</td><td>tacgtaggga</td><td>gcaagcgttg</td><td>tccggaatta</td><td>ctgggtgtaa</td><td>540</td>

<td>agggagcgta</td><td>ggcgggagga</td><td>taaagttgaa</td><td>tgtgaaatct</td><td>atgggctcaa</td><td>cccatagctg</td><td>600</td>

<td>cgttcaaaac</td><td>tgttcttctt</td><td>gagtgaagta</td><td>gaggcaggcg</td><td>gaattcctag</td><td>tgtagcggtg</td><td>660</td>

<td>aaatgcgtag</td><td>atattaggag</td><td>gaacaccagt</td><td>ggcgaaagcg</td><td>gcctgctggg</td><td>cttttactga</td><td>720</td>

<td>cgctgaggct</td><td>cgaaagcgtg</td><td>ggtagcaaac</td><td>agaattagat</td><td>taccctgtta</td><td>ttcacggcgg</td><td>780</td>

<td>taaacgatga</td><td>ttactaggtt</td><td>tgggttgacc</td><td>tgacccccat</td><td>tcgtgccgga</td><td>agtaacacca</td><td>840</td>

<td>taaagtaatc</td><td>cacctggggg</td><td>agtacggccg</td><td>ccaggttgaa</td><td>acttcaaaag</td><td>gaattgacgg</td><td>900</td>

<td>gggcccgcac</td><td>aagcagtgga</td><td>ggtatgtggt</td><td>ttaatttcga</td><td>cgcaaacgcg</td><td>aagaacctta</td><td>960</td>

<td>ccagggtctt</td><td>gacatcgagt</td><td>gacggacata</td><td>gagatatgtc</td><td>tttcctttcg</td><td>ggacacgaag</td><td>1020</td>

<td>acaggtggtg</td><td>catggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>gttgggttaa</td><td>gtcccgcaac</td><td>1080</td>

<td>gagcgcaacc</td><td>cttaccatta</td><td>gttgctacgc</td><td>aagagcactc</td><td>tgatgggact</td><td>gccgttgaca</td><td>1140</td>

<td>aaacggagga</td><td>aggtggggat</td><td>gacgtcaaat</td><td>catcatgccc</td><td>cttatgacct</td><td>gggcgacaca</td><td>1200</td>

<td>cgtactacaa</td><td>tggcggtcaa</td><td>cagagggagg</td><td>caaagccgcg</td><td>aggcagagca</td><td>aacccctaaa</td><td>1260</td>

<td>agccgtctca</td><td>gttcggattg</td><td>caggctgcaa</td><td>ctcgcctgca</td><td>tgaagtcgga</td><td>attgctagta</td><td>1320</td>

<td>atcgcggatc</td><td>agcatgccgc</td><td>ggtgaatacg</td><td>ttcccgggcc</td><td>ttgtacacac</td><td>cgcccgtcac</td><td>1380</td>

<td>accatgagag</td><td>ccggtaacac</td><td>ccgaagtcaa</td><td>tagtctaacc</td><td>gcaaggagga</td><td>cattgccgaa</td><td>1440</td>

<td>ggtgggattg</td><td>gtaattgggg</td><td>tgaagtcgta</td><td>acaaggtagc</td><td>cgt</td><td></td><td>1483</td>

<210> 40 <211> 1511 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1)..(1511) <223> Sekwencja genu kodującego 16S rRNA 20 szczepu Clostridium <400> 40

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggctcatat</td><td>tgaaacctag</td><td>tgatgtatga</td><td>gttagtggcg</td><td>gacgggtgag</td><td>taacgcgtgg</td><td>120</td>

<td>agaacctgcc</td><td>gtatactggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>gcataagcgc</td><td>180</td>

<td>acagcttcgc</td><td>atgaaacagt</td><td>gtgaaaaact</td><td>ccggtggtat</td><td>acgatggatc</td><td>cgcgtctgat</td><td>240</td>

<td>tagctggttg</td><td>gcggggtaac</td><td>agcccaccaa</td><td>ggcgacgatc</td><td>agtagccggc</td><td>ctgagagggt</td><td>300</td>

<td>gaacggccac</td><td>attgggactg</td><td>agacacggcc</td><td>caaactccta</td><td>cgggaggcag</td><td>cagtggggaa</td><td>360</td>

<td>tattgcacaa</td><td>tgggggaaac</td><td>cctgatgcag</td><td>cgacgccgcg</td><td>tgagtgaaga</td><td>agtatttcgg</td><td>420</td>

<td>tatgtaaagc</td><td>tctatcagca</td><td>gggaagaaat</td><td>actgacctta</td><td>cggtcagcag</td><td>acggtacctg</td><td>480</td>

<td>actaagaagc</td><td>cccggctaac</td><td>tacgtgccag</td><td>cagccgcggt</td><td>aatacgtagg</td><td>ggcaagcgtt</td><td>540</td>

<td>atccggattt</td><td>actgggtgta</td><td>aagggagcgt</td><td>agacggcagc</td><td>gcaagtctga</td><td>gtgaaatccc</td><td>600</td>

<td>atggcttaac</td><td>catggaactg</td><td>ctttggaaac</td><td>tgtgcagctg</td><td>gagtgcagga</td><td>gaggtaaagc</td><td>660</td>

<td>ggaattccta</td><td>gtgtagcggg</td><td>tgaaatgcgt</td><td>agatatagga</td><td>ggaacaacag</td><td>tggcggaagg</td><td>720</td>

<td>cggctactgg</td><td>gactgtaact</td><td>gacgttgagg</td><td>ctcgaaagcg</td><td>tggggagcaa</td><td>acaggattag</td><td>780</td>

<td>ataccctggt</td><td>agtcacgccg</td><td>taaacgatga</td><td>ttactaggtg</td><td>ttgggggacc</td><td>ataggtcttc</td><td>840</td>

<td>ggtgccggcg</td><td>caaacgcaat</td><td>taagtaatcc</td><td>acctggggga</td><td>gtacgttcgc</td><td>aagaatgaaa</td><td>900</td>

<td>ctcaaaggaa</td><td>ttgacgggga</td><td>cccgcacaaa</td><td>gcggtggagc</td><td>atgtggttta</td><td>attcgaaagc</td><td>960</td>

<td>aaacgcgaag</td><td>aaaccttacc</td><td>tggtcttgac</td><td>atcccgatga</td><td>cgagtgagca</td><td>aagtcacttt</td><td>1020</td>

<td>cccttcgggg</td><td>caattggaga</td><td>caggtggtgc</td><td>atgggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>1080</td>

<td>gttgggttaa</td><td>gtcccgcaac</td><td>gagcgcaacc</td><td>cctatttcca</td><td>gtagccagca</td><td>ggtagagctg</td><td>1140</td>

<td>ggcactctgg</td><td>agagactgcc</td><td>cgggataacc</td><td>gggaggaagg</td><td>cggggatgac</td><td>gtcaaatcat</td><td>1200</td>

<td>catgcccctt</td><td>atgatcaggg</td><td>ctacacacgt</td><td>gctacaatgg</td><td>cgtaaacaaa</td><td>gggaagcgag</td><td>1260</td>

<td>acggtgacgt</td><td>taagcaaatc</td><td>ccaaaaataa</td><td>cgtcccagtt</td><td>cggattgtag</td><td>tctgcaactc</td><td>1320</td>

<td>gattacatga</td><td>agctggaatc</td><td>gctagtaatc</td><td>gcgaatcaga</td><td>atgtcgcggt</td><td>gaatacgttc</td><td>1380</td>

<td>ccgggtcttg</td><td>tacacaccgc</td><td>ccgtcacacc</td><td>atgggagtcg</td><td>gaaatgcccg</td><td>aagtcagtga</td><td>1440</td>

<td>cctaaccgaa</td><td>aggaaggagc</td><td>tgccgaaggt</td><td>ggagccggta</td><td>actggggtga</td><td>agtagataac</td><td>1500</td>

<td>aaggtagccg</td><td>t .</td><td></td><td></td><td></td><td></td><td>1511</td>

<210> 41 <211> 1495 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1495) <223> Sekwencja genu kodującego 16S rRNA 21 szczepu Clostridium <400> 41

<td>agagtttgat</td><td>cctgcgctca</td><td>ggacgaacgc</td><td>tggcggcgcg</td><td>cctaacacat</td><td>gcaagtcgaa</td><td>60</td>

<td>cgggactatt</td><td>ttgggagaag</td><td>ttttcggatg</td><td>gatctcggga</td><td>tagtttagtg</td><td>gcggacgggt</td><td>120</td>

<td>gagtaacgcg</td><td>tgggcaacct</td><td>gccttacaca</td><td>gggggataac</td><td>aattagaaat</td><td>gattgctaat</td><td>180</td>

<td>accgcataag</td><td>accacggtac</td><td>tgcatggtac</td><td>agtggtaaaa</td><td>actgaggtgg</td><td>tgtaagatgg</td><td>240</td>

<td>gcccgcgtct</td><td>gattaggtag</td><td>ttggtggggt</td><td>agaagcctac</td><td>caagccgacg</td><td>atcagtagcc</td><td>300</td>

<td>gacctgagag</td><td>ggcgaccggc</td><td>cacattggga</td><td>ctgagacacg</td><td>gcccaaactc</td><td>ctacgggagg</td><td>360</td>

<td>cagcagtggg</td><td>gaatattgca</td><td>caatggggga</td><td>aaccctgatg</td><td>cagcgacgcc</td><td>gcgtgagtga</td><td>420</td>

<td>ggaagtattt</td><td>cggtatgtaa</td><td>agctctatca</td><td>gcagggaaga</td><td>aaatgacggt</td><td>acctgactaa</td><td>480</td>

<td>gaagccccgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtagggggca</td><td>agcgttatcc</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gagcgtagac</td><td>ggacttgcaa</td><td>gtctgatgtg</td><td>aaaatccggg</td><td>600</td>

<td>ggcccaaccc</td><td>cggaactgca</td><td>ttggaaactg</td><td>tatatctaga</td><td>gtgtcggaga</td><td>ggcaagtgga</td><td>660</td>

<td>atttcctggt</td><td>gtagcggtga</td><td>aatgcgtaga</td><td>tatcagagga</td><td>acaccagtgg</td><td>cgaaggcgct</td><td>720</td>

<td>tgcctgacga</td><td>tgactgacgt</td><td>tgaagctcga</td><td>aaagcgtggg</td><td>tagcaaacag</td><td>aattagatac</td><td>780</td>

<td>cctggtaagt</td><td>caacccggta</td><td>aacgatgatt</td><td>actaggtttt</td><td>ggttggactg</td><td>accccatccg</td><td>840</td>

<td>tgccggagta</td><td>acaccaataa</td><td>gttatccaac</td><td>ctgggaagta</td><td>cggccggcag</td><td>gttgaaactc</td><td>900</td>

<td>aaaaggaaat</td><td>gacgggggcc</td><td>cgcacaagca</td><td>gttgaagtat</td><td>gtgggttaat</td><td>tcgacgcaaa</td><td>960</td>

<td>cgcgaagaac</td><td>cttaccaggt</td><td>cttgacatcg</td><td>agtgacggac</td><td>atagagatat</td><td>gtctttcctt</td><td>1020</td>

<td>cgggacacga</td><td>agacaggtgg</td><td>tgcatggttg</td><td>tcgtcagctc</td><td>gtgtcgtgag</td><td>atgttgggtt</td><td>1080</td>

<td>aagtcccgca</td><td>acgagcgcaa</td><td>cccttaccat</td><td>tagttgctac</td><td>gcaagagcac</td><td>tctgatggga</td><td>1140</td>

<td>ctgccgttga</td><td>caaaacggag</td><td>gaaggtgggg</td><td>atgacgtcaa</td><td>atcatcatgc</td><td>cccttatgac</td><td>1200</td>

<td>ctgggcgaca</td><td>cacgtactac</td><td>aatggcggtc</td><td>aacagaggga</td><td>ggcaaagccg</td><td>cgaggcagag</td><td>1260</td>

<td>caaaccccta</td><td>aaagccgtct</td><td>cagttcggat</td><td>tgcaggctgc</td><td>aactcgcctg</td><td>catgaagtcg</td><td>1320</td>

<td>gaattgctag</td><td>taatcgcgga</td><td>tcagcatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>ccttgtacac</td><td>1380</td>

<td>accgcccgtc</td><td>acaccatgag</td><td>agccggtaac</td><td>acccgaagtc</td><td>aatagtctaa</td><td>ccgcaaggag</td><td>1440</td>

<td>gacattgccg</td><td>aaggtgggat</td><td>tggtaattgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td>1495</td>

<210> 42 <211> 1491 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1491) <223> Sekwencja genu kodującego 16S rRNA 22 szczepu Clostridium <400> 42

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gagaatccag</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatctggag</td><td>gaaagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtga</td><td>gcaatctgcc</td><td>ttggagtggg</td><td>gaataacggt</td><td>tggaaacagc</td><td>cgctaatacc</td><td>180</td>

<td>gcatgatgcg</td><td>tctgggaggc</td><td>atctctctgg</td><td>acgccaaaga</td><td>tttatcgctc</td><td>tgagatgagc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagcttgtt</td><td>ggcggggtaa</td><td>aggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgcaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttctg</td><td>agggacgaag</td><td>aaagtgacgg</td><td>tacctcagga</td><td>480</td>

<td>ataagccacg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtaggtggc</td><td>aagcgttatc</td><td>540</td>

<td>cggatttatt</td><td>gggtgtaaag</td><td>ggcgtgtagg</td><td>cgggaaagca</td><td>agtcagatgt</td><td>gaaaactcag</td><td>600</td>

<td>ggctcaaccc</td><td>tgagcctgca</td><td>tttgaaactg</td><td>tttttcttga</td><td>gtgctggaga</td><td>ggcaatcgga</td><td>660</td>

<td>attccgtgtg</td><td>tagcggtgaa</td><td>atgcgtagat</td><td>atacggagga</td><td>caccagtggc</td><td>gaagcggatt</td><td>720</td>

<td>gctggacagt</td><td>aactgacgct</td><td>gaggcgcgaa</td><td>gcgtggggag</td><td>caaacaggat</td><td>tagataccct</td><td>780</td>

<td>ggtagtccac</td><td>gccgtaaacg</td><td>atggatacta</td><td>ggtgtggggg</td><td>gactgacccc</td><td>ctccgtgccg</td><td>840</td>

<td>cagctaacgc</td><td>aataagtatc</td><td>ccacctgggg</td><td>agtacgatcg</td><td>caaggttgaa</td><td>actcaaagga</td><td>900</td>

<td>attgacgggg</td><td>gcccgcacaa</td><td>gcggtggagt</td><td>atgtggttta</td><td>attcgaagca</td><td>acgcgaagaa</td><td>960</td>

<td>ccttaccagg</td><td>gcttgacatc</td><td>ctgctaacga</td><td>accagagatg</td><td>gattaggtgc</td><td>ccttcgggga</td><td>1020</td>

<td>aagcagagac</td><td>aggtggtgca</td><td>tggttgtcgt</td><td>cagctcgtgt</td><td>cgtgagatgt</td><td>tgggttaagt</td><td>1080</td>

<td>cccgcaacga</td><td>gcgcaaccct</td><td>tattgttagt</td><td>tgctacgcaa</td><td>gagcactcta</td><td>gcgagactgc</td><td>1140</td>

<td>cgttgacaaa</td><td>acggaggaag</td><td>gtggggacga</td><td>cgtcaaatca</td><td>tcatgcccct</td><td>tacgtcctgg</td><td>1200</td>

<td>gccacacacg</td><td>tactacaatg</td><td>gcggccaaca</td><td>aagagaggca</td><td>agaccgcgag</td><td>gtggagcaaa</td><td>1260</td>

<td>tctcaaaaag</td><td>ccgtcccagt</td><td>tcggatcgca</td><td>ggctgcaacc</td><td>cgcctgcgtg</td><td>aagttggaat</td><td>1320</td>

<td>cgctagtaat</td><td>cgcggatcag</td><td>catgccgcgg</td><td>tgaatacgtt</td><td>cccgggcctt</td><td>gtacacaccg</td><td>1380</td>

<td>cccgtcacac</td><td>catgagagtc</td><td>gggaacaccc</td><td>gaagtccgta</td><td>gcctaaccgc</td><td>aaggggggcg</td><td>1440</td>

<td>cggccgaagg</td><td>tgggttcgat</td><td>aattggggtg</td><td>aagtcgtaac</td><td>aaggtagccg</td><td>t</td><td>1491</td>

<210> 43 <211> 1495 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1495) <223> Sekwencja genu kodującego 16S rRNA 23 szczepu Clostridium <400> 43

<td>agagtttgat</td><td>cctgtgcctc</td><td>aggatgaacg</td><td>ctggcggcgt</td><td>gcttaacaca</td><td>tgcaagtcga</td><td>60</td>

<td>acgagaacca</td><td>acggattgag</td><td>gattcgtcca</td><td>aatgaagttg</td><td>gggaaagtgg</td><td>cggacgggtg</td><td>120</td>

<td>agtaacgcgt</td><td>gagcaatctg</td><td>ccttggagtg</td><td>gggaataacg</td><td>gttggaaaca</td><td>gccgctaata</td><td>180</td>

<td>ccgcatgatg</td><td>cgtctgggag</td><td>gcatctctct</td><td>ggacgccaaa</td><td>gatttatcgc</td><td>tctgagatga</td><td>240</td>

<td>gctcgcgtct</td><td>gattagctag</td><td>ttggcggggc</td><td>aacggcccac</td><td>caaggcgacg</td><td>atcagtagcc</td><td>300</td>

<td>ggactgagag</td><td>gttggccggc</td><td>cacattggga</td><td>ctgagacacg</td><td>gcccagactc</td><td>ctacgggagg</td><td>360</td>

<td>cagcagtggg</td><td>gaatattggg</td><td>caatgggcgc</td><td>aagcctgacc</td><td>cagcaacgcc</td><td>gcgtgaagga</td><td>420</td>

<td>agaaggcttt</td><td>cgggttgtaa</td><td>acttctttta</td><td>agggggacga</td><td>acaaatgacg</td><td>gtaccccttg</td><td>480</td>

<td>aataagccac</td><td>ggctaactac</td><td>gtgccagcag</td><td>ccgcggtaat</td><td>acgtaggtgg</td><td>caagcgttat</td><td>540</td>

<td>ccggatttat</td><td>tgggtgtaaa</td><td>gggcgtgtag</td><td>gcgggaatgc</td><td>aagtcagatg</td><td>tgaaaactat</td><td>600</td>

<td>gggctcaacc</td><td>catagcctgc</td><td>atttgaaact</td><td>gtatttcttg</td><td>agtgctggag</td><td>aggcaatcgg</td><td>660</td>

<td>aattccgtgt</td><td>gtagcggtga</td><td>aatgcgtaga</td><td>tatacggagg</td><td>aacaccagtg</td><td>gcgaaggcgg</td><td>720</td>

<td>attgctggac</td><td>agtaactgac</td><td>gctgaggcgc</td><td>gaaagcgtgg</td><td>ggagcaaaca</td><td>ggattagata</td><td>780</td>

<td>ccctggtagt</td><td>ccacgccgta</td><td>aacgatggat</td><td>actaagtgtg</td><td>gggggactga</td><td>ccccctccgt</td><td>840</td>

<td>gccgcagcta</td><td>acgcaataag</td><td>tatcccacct</td><td>ggggagtacg</td><td>atcgcaaggt</td><td>tgaaactcaa</td><td>900</td>

<td>aggaattgac</td><td>gggggcccgc</td><td>acaagcggtg</td><td>gagtatgtgg</td><td>tttaattcga</td><td>agcaacgcga</td><td>960</td>

<td>agaaccttac</td><td>cagggcttga</td><td>catcctgcta</td><td>acgaaccaga</td><td>gatggatcag</td><td>gtgcccttcg</td><td>1020</td>

<td>gggaaagcag</td><td>agacaggtgg</td><td>tgcatggttg</td><td>tcgtcagctc</td><td>gtgtcgtgag</td><td>atgttgggtt</td><td>1080</td>

<td>aagtcccgca</td><td>acgagcgcaa</td><td>cccctattgt</td><td>tagttgctac</td><td>gcaagagcac</td><td>tctagcgaga</td><td>1140</td>

<td>ctgccgttga</td><td>caaaacggag</td><td>gaaggtgggg</td><td>acgacgtcaa</td><td>atcatcatgc</td><td>cccttacgtc</td><td>1200</td>

<td>ctgggccaca</td><td>cacgtactac</td><td>aatggcggcc</td><td>aacaaagaga</td><td>ggcaagaccg</td><td>cgaggtggag</td><td>1260</td>

<td>caaatctcaa</td><td>aaagccgtcc</td><td>cagttcggat</td><td>cgcaggctgc</td><td>aacccgcctg</td><td>cgtgaagttg</td><td>1320</td>

<td>gaatcgctag</td><td>taatcgcgga</td><td>tcagcatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>ccttgtacac</td><td>1380</td>

<td>accgcccgtc</td><td>acaccatgag</td><td>agtcgggaac</td><td>acccgaagtc</td><td>cgtagcctga</td><td>ccgcaagggg</td><td>1440</td>

<td>ggcgcggccg</td><td>aaggtgggtt</td><td>cgataattgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td>1495</td>

<210> 44 <211> 1440 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1440) <223> Sekwencja genu kodującego 16S rRNA 24 szczepu Clostridium <400> 44

<td>agagtttgat cctggctcag gacgaacgct ggcggcacgc ctaacacatg caagtcgaac</td><td>60</td>

<td>ggagtgaaga tgctcgcatc tgaacttagt ggcggacggg tgagtaacac gtgagcaacc</td><td>120</td>

<td>tgcctttcag agggggataa cgtttggaaa cgaacgctaa taccgcataa aatatcggag</td><td>180</td>

<td>tcgcatggca ctgatatcaa aggagtaatc ćgctgaaaga tgggctcgcg tccgattagg</td><td>240</td>

<td>cagttggcgg ggtaacggcc caccaaaccg acaatcggta gccggactga gaggttgaac</td><td>300</td>

<td>ggccacattg ggactgagac acggcccaga ctcctacggg aggcagcagt gggggatatt</td><td>360</td>

<td>gcacaatggg ggaaaccctg atgcagcgat gccgcgtgaa tgaagacggc cttcgggttg</td><td>420</td>

<td>taaagttctg tcgcagggga cgaaaatgac ggtaccctgc aagaaagctc cggctaacta</td><td>480</td>

<td>cgtgccagca gccgcggtaa tacgtaggga gcaagcgttg tccggaatta ctgggtgtaa</td><td>540</td>

<td>agggagcgta ggcgggagga taagttgaat gtgaaatcta tgggctcaac ccatagttgc</td><td>600</td>

<td>gttcaaaact gttcttcttg agtgaagtag aggcaggcgg aattcctagt gtagcggtga</td><td>660</td>

<td>aatgcgtaga tattagagga acaccagtgg cgaagcggcc tgctgggctt ttactgacgc</td><td>720</td>

<td>tgagctcgaa agcgtgggta gcaacaggat tagataccct ggtagtccac gcggtaaacg</td><td>780</td>

<td>atgattacta gtgtgggtgg actgacccat ccatgccgga gttaacacaa tagtaatcca</td><td>840</td>

<td>cctggggagt acgcgcagtg aactcaaagg attgacgggg cccgcacaag cagtgagtat</td><td>900</td>

<td>gtggtttatt cgacgcacgc gagactacag tcttgacatc gatgacggac tagagatatg</td><td>960</td>

<td>tctttctcgg acacgaagac aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt</td><td>1020</td>

<td>tgggttaagt cccgcaacga gcgcaaccct taccattagt tgttacgcaa gagcactcta</td><td>1080</td>

<td>atgggactgc cgttgacaaa acggaggaag gtggggatga cgtcaaatca tcatgcccct</td><td>1140</td>

<td>tatgacctgg gcgacacacg tactacaatg gcggtcaaca gagggaggca aagccgcgag</td><td>1200</td>

<td>gcagagcaaa cccctaaaag ccgtctcagt tcggattgca ggctgcaact cgcctgcatg</td><td>1260</td>

<td>aagtcggaat tgctagtaat cgcggatcag catgccgcgg tgaatacgtt cccgggcctt</td><td>1320</td>

<td>gtacacaccg cccgtcacac catgagagcc ggtaacaccc gaagtcaata gtctaaccgc</td><td>1380</td>

<td>aaggaggaca ttgccgaagg tgggatggta attggggtga agtagtaaca aggtagccgt</td><td>1440</td>

<210> 45 <211> 1495 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1495) <223> Sekwencja genu kodującego 16S rRNA 25 szczepu Clostridium <400> 45

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gagaaccatt</td><td>ggatcgagga</td><td>ttcgtccaag</td><td>tgaaggtggg</td><td>gaaagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtga</td><td>gcaatctgcc</td><td>ttggagtggg</td><td>gaataacggc</td><td>tggaaacagc</td><td>cgctaatacc</td><td>180</td>

<td>gcatgataca</td><td>gctgggaggc</td><td>atctccctgg</td><td>ctgtcaaaga</td><td>tttatcgctc</td><td>tgagatgagc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggcggggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgcaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttgtc</td><td>agggacgaag</td><td>caagtgacgg</td><td>tacctgacga</td><td>480</td>

<td>ataagccacg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtagtggca</td><td>agcgttatcc</td><td>540</td>

<td>ggatttattg</td><td>gggtgtaaag</td><td>ggcgtgtagg</td><td>cgggaatgca</td><td>agtcagatgt</td><td>gaaaactatg</td><td>600</td>

<td>gggctcaacc</td><td>catagcctgc</td><td>atttgaaact</td><td>gtatttcttg</td><td>agtgctggag</td><td>aggcaatcga</td><td>660</td>

<td>attccgtgtg</td><td>tagcgggtga</td><td>aatgcgtaga</td><td>tatacggagg</td><td>aacaccagtg</td><td>gcgaagcgga</td><td>720</td>

<td>ttgctggaca</td><td>agtaactgac</td><td>gctgaggcgc</td><td>gaaagcgtgg</td><td>ggagcaaaca</td><td>ggattagata</td><td>780</td>

<td>ccctggtagt</td><td>ccacgccgta</td><td>aacgatggat</td><td>actaggtgtg</td><td>gggggactga</td><td>ccccctccgt</td><td>840</td>

<td>gccgcagcta</td><td>acgcaataag</td><td>tatcccacct</td><td>ggggagtacg</td><td>atcgcaaggt</td><td>tgaaactcaa</td><td>900</td>

<td>aggaattgac</td><td>gggggcccgc</td><td>acaagcggtg</td><td>gagtatgtgg</td><td>tttaattcga</td><td>cgcaacgcga</td><td>960</td>

<td>agaaccttac</td><td>cagggcttga</td><td>catcctacta</td><td>acgaaccaga</td><td>gatggattag</td><td>gtgcccttcg</td><td>1020</td>

<td>gggaaagtag</td><td>agacaggtgg</td><td>tgcatggttg</td><td>tcgtcagctc</td><td>gtgtcgtgag</td><td>atgttgggtt</td><td>1080</td>

<td>aagtcccgca</td><td>acgagcgcaa</td><td>cccctattgt</td><td>tagttgctac</td><td>gcaagagcac</td><td>tctagcgaga</td><td>1140</td>

<td>ctgccgttga</td><td>caaaacggag</td><td>gaaggtgggg</td><td>acgacgtcaa</td><td>atcatcatgc</td><td>cccttacgtc</td><td>1200</td>

<td>ctgggccaca</td><td>cacgtactac</td><td>aatggcggcc</td><td>aacaaagaga</td><td>ggcaaagccg</td><td>cgaggtggag</td><td>1260</td>

<td>caaatctcaa</td><td>aaagccgtcc</td><td>cagttcggat</td><td>cgcaggctgc</td><td>aacccgcętg</td><td>cgtgaagttg</td><td>1320</td>

<td>gaatcgctag</td><td>taatcgcgga</td><td>tcagcatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>ccttgtacac</td><td>1380</td>

<td>accgcccgtc</td><td>acaccatgag</td><td>agtcgggaac</td><td>acccgaagtc</td><td>cgtagcctaa</td><td>ccgcaagggg</td><td>1440</td>

<td>ggcgcggccg</td><td>aaggtgggtt</td><td>cgataattgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td>1495</td>

<210> 46 <211> 1495 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1495) <223> Sekwencja genu kodującego 16S rRNA 26 szczepu Clostridium

<td>&lt;400&gt; 46 agagtttgat</td><td>catggctcag</td><td>gacgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatgggaat</td><td>gcttagtggc</td><td>ggactggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>cttccagagg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcatgatgc</td><td>gttggagccg</td><td>catgactccg</td><td>acgtcaaaga</td><td>tttatcgctg</td><td>gaagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggtgaggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atggacgcaa</td><td>gtctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>ggggaagagc</td><td>agaagacggt</td><td>accccttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttgtcc</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gcgtgcagcc</td><td>ggagagacaa</td><td>gtcagatgtg</td><td>aaatccacgg</td><td>600</td>

<td>gctcaacccg</td><td>tgaactgcat</td><td>ttgaaactgt</td><td>ttcccttgag</td><td>tgtcggagag</td><td>ggtaatcgga</td><td>660</td>

<td>attcctttgt</td><td>gtagcggtga</td><td>aatgcgtaga</td><td>tataagaaga</td><td>acaccagtgg</td><td>cgaaggcgga</td><td>720</td>

<td>ttactggacg</td><td>ataactgacg</td><td>gtgaggcgcg</td><td>aaagcgtggg</td><td>ggagcaacag</td><td>attaaatacc</td><td>780</td>

<td>ctggtagtcc</td><td>acgctgttaa</td><td>cgatcgatac</td><td>taggtgtgcc</td><td>gggactgacc</td><td>ccctgcgtgc</td><td>840</td>

<td>ccggagttaa</td><td>ccacaataag</td><td>tatcgcacct</td><td>ggggagtacg</td><td>atcgcaaggt</td><td>gaacttcaaa</td><td>900</td>

<td>ggaattgacg</td><td>ggggcccgcc</td><td>ccaagccgtg</td><td>gattatgtgg</td><td>ttaattcgaa</td><td>gcaacgcgaa</td><td>960</td>

<td>gaacctaccc</td><td>agggcttgac</td><td>atcctgctaa</td><td>cgaagtagag</td><td>atacattagg</td><td>tgccctttcg</td><td>1020</td>

<td>gggaaagcag</td><td>agacaggtgg</td><td>tgcatggttg</td><td>tcgtcagctc</td><td>gtgtcgtgag</td><td>atgttgggtt</td><td>1080</td>

<td>aagtcccgca</td><td>acgagcgcaa</td><td>cccctattgt</td><td>tagttgctac</td><td>gcaagagcac</td><td>tctagcgaga</td><td>1140</td>

<td>ctgccgttga</td><td>caaaacggag</td><td>gaaggcgggg</td><td>acgacgtcaa</td><td>atcatcatgc</td><td>cccttatgtc</td><td>1200</td>

<td>ctgggctaca</td><td>cacgtaatac</td><td>aatggcggtt</td><td>aacaaaggga</td><td>tgcaaagccg</td><td>cgaggcagag</td><td>1260</td>

<td>cgaaccccaa</td><td>aaagccgtcc</td><td>cagttcggat</td><td>cgcaggctgc</td><td>aacccgcctg</td><td>cg.tgaagtcg</td><td>1320</td>

<td>gaatcgctag</td><td>taatcgcgga</td><td>tcagcatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>ccttgtacac</td><td>1380</td>

<td>accgcccgtc</td><td>acaccatgag</td><td>agtcgggaac</td><td>acccgaagtc</td><td>cgtagcctaa</td><td>ccgcaaggag</td><td>1440</td>

<td>ggcgcggccg</td><td>aaggtgggtt</td><td>cgataattgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td>1495</td>

<210> 47 <211> 1509 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1509) <223> Sekwencja genu kodującego 16S rRNA 27 szczepu Clostridium

-77<400> 47

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagtacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>tgatgggact</td><td>acttagtggc</td><td>ggacgggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>agtaacctgc</td><td>cttggagtgg</td><td>ggaataacag</td><td>ctggaaacag</td><td>ctgctaatac</td><td>180</td>

<td>cgcataatat</td><td>gtctgtgtcg</td><td>catggcactg</td><td>gacatcaaag</td><td>atttatcgct</td><td>ctgagatgga</td><td>240</td>

<td>ctcgcgtctg</td><td>attagctagt</td><td>tggcggggta</td><td>acggcccacc</td><td>aaggcgacga</td><td>tcagtagccg</td><td>300</td>

<td>gactgagagg</td><td>ttggccggcc</td><td>acattgggac</td><td>tgagacacgg</td><td>cccagactcc</td><td>tacgggaggc</td><td>360</td>

<td>agcagtgggg</td><td>aatattgggc</td><td>aatgggcgca</td><td>agcctgaccc</td><td>agcaacgccg</td><td>cgtgaaggaa</td><td>420</td>

<td>gaaggctttc</td><td>gggttgtaaa</td><td>cttcttttaa</td><td>gggggaagag</td><td>cagaagacgg</td><td>taccccttga</td><td>480</td>

<td>ataagccacg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtaggtggc</td><td>aagcgttgtc</td><td>540</td>

<td>cggatttact</td><td>gggtgtaaag</td><td>ggcgtgcagc</td><td>cggagagaca</td><td>agtcagatgt</td><td>gaaatccacg</td><td>600</td>

<td>ggctcaaccc</td><td>gtgaactgca</td><td>tttgaaactg</td><td>tttcctggag</td><td>ttcggagggt</td><td>atggaattct</td><td>660</td>

<td>tgttagcggt</td><td>gaaatgctgt</td><td>agatatggga</td><td>gaaccaccag</td><td>tgcgaggggg</td><td>cttccgggac</td><td>720</td>

<td>tgtacttgac</td><td>tgtagaggtc</td><td>tcaaagctgg</td><td>gggagcaccg</td><td>aggaatgaga</td><td>taccgtgata</td><td>780</td>

<td>gtcccacgcg</td><td>gtaacggatg</td><td>attactaggt</td><td>gttgggggga</td><td>cccaggctct</td><td>ttcggtgccg</td><td>840</td>

<td>ggcgcaaacc</td><td>ctttaggaat</td><td>tccacctggg</td><td>gaattacgtt</td><td>tggcaagaaa</td><td>ggaacttcaa</td><td>900</td>

<td>agaaattgaa</td><td>cgggggaccc</td><td>ccccaaccgg</td><td>tggaggcatg</td><td>gtgttttatt</td><td>tcggaggaac</td><td>960</td>

<td>gggaagaacc</td><td>tttaccttgt</td><td>tctgaccttc</td><td>cggatgacga</td><td>agtgagcaaa</td><td>gtcaacttcc</td><td>1020</td>

<td>cttcggggcc</td><td>atggaggaca</td><td>ggtggtggca</td><td>tggttggtcg</td><td>tcagctcgtg</td><td>tcgtgagatg</td><td>1080</td>

<td>ttgggttaag</td><td>tcccgcaacg</td><td>agcgcaaccc</td><td>ctatttccag</td><td>tagccagcag</td><td>gtagagctgg</td><td>1140</td>

<td>gcactctgga</td><td>gagactgccc</td><td>gggataaccg</td><td>ggaggaaggc</td><td>ggggatgacg</td><td>tcaaatcatc</td><td>1200</td>

<td>atgcccctta</td><td>tgatcagggc</td><td>tacacacgtg</td><td>ctacaatggc</td><td>gtaaacaaag</td><td>ggaagcgaga</td><td>1260</td>

<td>cggtgacgtt</td><td>aagcaaatcc.</td><td>caaaaataac</td><td>gtcccagttc</td><td>ggattgtagt</td><td>ctgcaactcg</td><td>1320</td>

<td>actacatgaa</td><td>gctggaatcg</td><td>ctagtaatcg</td><td>cgaatcagaa</td><td>tgtcgcggtg</td><td>aatacgttcc</td><td>1380</td>

<td>cgggtcttgt</td><td>acacaccgcc</td><td>cgtcacacca</td><td>tgggagtcgg</td><td>aaatgcccga</td><td>agtcagtgac</td><td>1440</td>

<td>ctaaccgaaa</td><td>ggaaggagct</td><td>gccgaaggtg</td><td>gagccggtaa</td><td>ctggggtgaa</td><td>gtcgtaacaa</td><td>1500</td>

<td>ggtagccgt</td><td></td><td></td><td></td><td></td><td></td><td>1509</td>

<210> 48 <211> 1583 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1583) <223> Sekwencja genu kodującego 16S rRNA 28 szczepu Clostridium

-78<400> 48

<td>agagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcgcgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcttata</td><td>tttcagaagt</td><td>tttcggatgg</td><td>acgagagata</td><td>agcttagtgg</td><td>cggacgggtg</td><td>120</td>

<td>agtaacacgt</td><td>gagcaacctg</td><td>cctttcagag</td><td>ggggataaca</td><td>gttggaaacg</td><td>actgctaata</td><td>180</td>

<td>ccgcataacg</td><td>ctgcgatggg</td><td>gcatcccgat</td><td>gcagccaaag</td><td>gagcaatccg</td><td>ctgaaagatg</td><td>240</td>

<td>ggctcgcggc</td><td>cgattagcta</td><td>gttggtgggg</td><td>caacggccca</td><td>ccaaggcgac</td><td>gatcggtagc</td><td>300</td>

<td>cggactgaga</td><td>ggttgatcgg</td><td>ccacattggg</td><td>actgagacac</td><td>ggcccagact</td><td>cctacgggag</td><td>360</td>

<td>gcagcagtgg</td><td>gggatattgc</td><td>acaatggagg</td><td>aaactctgat</td><td>gcagcgacgc</td><td>cgcgtgaggg</td><td>420</td>

<td>aagacggtct</td><td>tcggattgta</td><td>aacctctgtc</td><td>tttggggaag</td><td>aaaatgacgg</td><td>tacccaaaga</td><td>480</td>

<td>ggaagctccg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtaggggag</td><td>cgagcgttgt</td><td>540</td>

<td>ccggaattac</td><td>tgggtgtaaa</td><td>gggagcgtag</td><td>cgggcgagaa</td><td>agttgaatgt</td><td>taaatctacc</td><td>600</td>

<td>ggcttaactg</td><td>gtagctgcgt</td><td>ccaaaacttc</td><td>ttggtcttga</td><td>gtgaaagtaa</td><td>gaggccaggg</td><td>660</td>

<td>cggaaattct</td><td>tagtgtaagc</td><td>gggtgaaaat</td><td>gcgttagata</td><td>ttagggagga</td><td>accaccaggt</td><td>720</td>

<td>gggcgaaggg</td><td>cggcttgctg</td><td>ggctttaact</td><td>ggacggctgg</td><td>aggcttggaa</td><td>aaggcgtggg</td><td>780</td>

<td>gagagcaaac</td><td>acagggaatt</td><td>aagtataccc</td><td>tggtatatgt</td><td>cacacgcttg</td><td>taaagagtat</td><td>840</td>

<td>gattaactta</td><td>gggtggtggg</td><td>gggaacttga</td><td>ccctttcgtg</td><td>tgcgcgcagg</td><td>ttaacacaca</td><td>900</td>

<td>tttagagtat</td><td>atccaacttg</td><td>gggagagtac</td><td>ggccggcaaa</td><td>gtttgaaact</td><td>tcaaaaggga</td><td>960</td>

<td>aattgagacc</td><td>ggggggcccg</td><td>gccaccaagc</td><td>acagtggaga</td><td>gtatggtggg</td><td>tttaatttcg</td><td>1020</td>

<td>agaagcaacc</td><td>ggcggaagag</td><td>aaactttacc</td><td>agtccttgac</td><td>atcggtggcg</td><td>gcataagccc</td><td>1080</td>

<td>tagagattag</td><td>gtgaagccct</td><td>tcgggggccc</td><td>caccagacag</td><td>gtggtgcatg</td><td>gttgtcgtca</td><td>1140</td>

<td>gctcgtgtcg</td><td>tgagatgttg</td><td>ggttaagtcc</td><td>ccgcaaacga</td><td>gcgcaaccct</td><td>tattattagt</td><td>1200</td>

<td>ttgctacgca</td><td>agagcactct</td><td>aatgagactg</td><td>ccgttgacaa</td><td>aacggaggaa</td><td>ggtggggatg</td><td>1260</td>

<td>acgtcaaatc</td><td>atcatgcccc</td><td>ttatgacctg</td><td>ggctacacac</td><td>gtactacaat</td><td>ggcactgaaa</td><td>1320</td>

<td>cagagggaag</td><td>cgacatcgcg</td><td>aggtgaagcg</td><td>aatcccaaaa</td><td>aagtgtccca</td><td>gttcggattg</td><td>1380</td>

<td>caggctgcaa</td><td>ctcgcctgca</td><td>tgaagtcgga</td><td>attgctagta</td><td>atcgcggatc</td><td>agcatgccgc</td><td>1440</td>

<td>ggtgaatacg</td><td>ttcccgggcc</td><td>ttgtacacac</td><td>cgcccgtcac</td><td>accatgggag</td><td>tcggtaacac</td><td>1500</td>

<td>ccgaagccag</td><td>tagcctaacc</td><td>gcaaggaggg</td><td>cgctgtcgaa</td><td>ggtgggattg</td><td>atgactgggg</td><td>1560</td>

<td>tgaagtcgta</td><td>acaaggtagc</td><td>cgt</td><td></td><td></td><td></td><td>1583</td>

<210> 49 <211> 1519 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1)..(1519) <223> Sekwencja genu kodującego 16S rRNA 29 szczepu Clostridium

-79<400> 49

<td>agagtttgat</td><td>catggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggtgtacgg</td><td>gaaggaaggc</td><td>ttcggccgga</td><td>aaacctgtgc</td><td>atgagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtgg</td><td>gcaacctggc</td><td>ctgtacaggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>180</td>

<td>gcataacggg</td><td>ggaagccgca</td><td>tggcttttcc</td><td>ctgaaaactc</td><td>cggtggtaca</td><td>ggatgggccc</td><td>240</td>

<td>gcgtctgatt</td><td>agccagttgg</td><td>cagggtaacg</td><td>gcctaccaaa</td><td>gcgacgatca</td><td>gtagccggcc</td><td>300</td>

<td>tgagagggcg</td><td>gacggccaca</td><td>ctgggactga</td><td>gacacggccc</td><td>agactcctac</td><td>gggaggcagc</td><td>360</td>

<td>agtgggggat</td><td>attgcacaat</td><td>ggggggaacc</td><td>ctgatgcagc</td><td>gacgccgcgt</td><td>gggtgaagaa</td><td>420</td>

<td>gcgcctcggc</td><td>gcgtaaagcc</td><td>ctgtcagcag</td><td>ggaaagaaaa</td><td>tgacggtacc</td><td>tgaagaagaa</td><td>480</td>

<td>gccccgggct</td><td>aactacgtgc</td><td>cagcagccgg</td><td>cggtaattac</td><td>gtaggggggc</td><td>aggcgttatc</td><td>540</td>

<td>cggatttact</td><td>gggtggtaaa</td><td>ggggggcgca</td><td>aacggcgatg</td><td>gcaggccagg</td><td>aatggaaagc</td><td>600</td>

<td>ccgggggccc</td><td>aaccccggga</td><td>cttgctcttg</td><td>ggaaactggc</td><td>ttggctggga</td><td>gtggcaggag</td><td>660</td>

<td>gggcaggcgg</td><td>aaattcctgg</td><td>tggtagcggt</td><td>ggaaaatggc</td><td>taaaaatcaa</td><td>gaagaaaaac</td><td>720</td>

<td>cggtggggaa</td><td>aggcggcctg</td><td>gtgggactgc</td><td>gaactgacgt</td><td>tgaaggcccg</td><td>aaagcgtggg</td><td>780</td>

<td>gaacaaacag</td><td>gatagattcc</td><td>ctggtagttc</td><td>cacgccgtaa</td><td>acgatgatta</td><td>ctaggtgtcg</td><td>840</td>

<td>gggagcagag</td><td>actgcccggt</td><td>gccgcagcca</td><td>acgcattaag</td><td>taatccacct</td><td>ggggagtacg</td><td>900</td>

<td>ttcgcaagaa</td><td>tgaaactcaa</td><td>aggaattgac</td><td>ggggacccgc</td><td>acaagcggtg</td><td>gagcatgtgg</td><td>960</td>

<td>tttaattcga</td><td>agcaacgcga</td><td>agaaccttac</td><td>ctggtcttga</td><td>catcccgatg</td><td>acgagtgagc</td><td>1020</td>

<td>aaagtcactt</td><td>tcccttcggg</td><td>gcattggaga</td><td>caggtggtgc</td><td>atggttgtcg</td><td>tcagctcgtg</td><td>1080</td>

<td>tcgtgagatg</td><td>ttgggttaag</td><td>tcccgcaacg</td><td>agcgcaaccc</td><td>ctatttccag</td><td>tagccagcag</td><td>1140</td>

<td>gtagagctgg</td><td>gcactctgga</td><td>gagactgccc</td><td>gggataaccg</td><td>ggaggaaggc</td><td>ggggatgacg</td><td>1200</td>

<td>tcaaatcatc</td><td>atgcccctta</td><td>tgatcagggc</td><td>tacacacgtg</td><td>ctacaatggc</td><td>gtaaacaaag</td><td>1260</td>

<td>ggaagcgaga</td><td>cggtgacgtt</td><td>aagcaaatcc</td><td>caaaaataac</td><td>gtcccagttc</td><td>ggattgtagt</td><td>1320</td>

<td>ctgcaactcg</td><td>actacatgaa</td><td>gctggaatcg</td><td>ctagtaatcg</td><td>cgaatcagaa</td><td>tgtcgcggtg</td><td>1380</td>

<td>aatacgttcc</td><td>cgggtcttgt</td><td>acacaccgcc</td><td>cgtcacacca</td><td>tgggagtcgg</td><td>aaatgcccga</td><td>1440</td>

<td>agtcagtgac</td><td>ctaaccgaaa</td><td>ggaaggagct</td><td>gccgaaggtg</td><td>gagccggtaa</td><td>ctggggtgaa</td><td>1500</td>

<td>gtcgtaacaa</td><td>ggtagccgt</td><td></td><td></td><td></td><td></td><td>1519</td>

<210> 50 <211> 1497 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1)..(1497) <223> Sekwencja genu kodującego 16S rRNA 30 szczepu Clostridium

-80<400> 50

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggggtatata</td><td>agcggaagtt</td><td>tacggatgga</td><td>aggttatata</td><td>cttagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtgg</td><td>gcaacctgcc</td><td>ccgtgccggg</td><td>ggataccgcc</td><td>tggaaacagg</td><td>cgctaatacc</td><td>180</td>

<td>gcataagcgc</td><td>atacagccgc</td><td>atgggtgtat</td><td>gcggaaagct</td><td>ccggcggcac</td><td>gggatgggcc</td><td>240</td>

<td>cgcgcccgat</td><td>tagccagttg</td><td>gcggggtaac</td><td>ggcccaccaa</td><td>agcgacgatc</td><td>ggtagccggc</td><td>300</td>

<td>ctgagagggc</td><td>ggacggccac</td><td>attgggactg</td><td>agacacggcc</td><td>caaactccta</td><td>cgggaggcag</td><td>360</td>

<td>cagtggggaa</td><td>tattgcacaa</td><td>tgggggaaac</td><td>cctgatgcag</td><td>caacgccgcg</td><td>tgggtgaagg</td><td>420</td>

<td>agcgtttcgg</td><td>cgcgtaaagc</td><td>cctgtcagcg</td><td>gggaagaaga</td><td>aagacggtac</td><td>ccgaccaaga</td><td>480</td>

<td>agccccggct</td><td>aactacgtgc</td><td>cagcagccgc</td><td>ggtaatacgt</td><td>agggggcgag</td><td>cgttatccgg</td><td>540</td>

<td>aattactggg</td><td>tgtaaaggga</td><td>gcgtagacgg</td><td>cgaggtaagc</td><td>ctgaagtgga</td><td>agcccgcggc</td><td> 600</td>

<td>ccaaccgcgg</td><td>aactgctttg</td><td>ggaactgttt</td><td>tgctggagta</td><td>tgggaggggt</td><td>aagcggaatt</td><td>660</td>

<td>cctggtgtag</td><td>cggtgaaatg</td><td>cgtagatatc</td><td>aggaggaaca</td><td>ccggtggcga</td><td>aggcggctta</td><td>720</td>

<td>ctggaccata</td><td>actgacgttg</td><td>aggctcgaaa</td><td>gcgtggggag</td><td>cgaacaggat</td><td>tagataccct</td><td>780</td>

<td>ggtagtccac</td><td>gcgtaaacga</td><td>tgattaccag</td><td>gtgtcgggtg</td><td>tcgaaggacg</td><td>gcccggtgcc</td><td>840</td>

<td>gcagcgaacg</td><td>cagtaagtaa</td><td>tccacctggg</td><td>gagtacgttc</td><td>gcaagaatga</td><td> aactcaaagg</td><td>900</td>

<td>aattgacggg</td><td>gacccgcaca</td><td>agcggtggag</td><td>catgtggttt</td><td>aattcgaagc</td><td>aacgcgaaga</td><td>960</td>

<td>accttacccg</td><td>gccttgacat</td><td>cccctggaca</td><td>gcatatgtaa</td><td>tgtatgtttc</td><td>cttcgggacc</td><td>1020</td>

<td>agggagacag</td><td>gtggtgcatg</td><td>gttgtcgtca</td><td>gctcgtgtcg</td><td>tgagatgttg</td><td>ggtcaagtcc</td><td>1080</td>

<td>cgcaacgagc</td><td>gcaacccctg</td><td>cccccagtag</td><td>ccagcattta</td><td>agatgggcac</td><td>tctgggggga</td><td>1140</td>

<td>ctgccgggga</td><td>taacccggag</td><td>gaaggcgggg</td><td>atgacgtcaa</td><td>atcatcatgc</td><td>cccttatggc</td><td>1200</td>

<td>cggggctaca</td><td>cacgtgctac</td><td>aatggcgtaa</td><td>acagagggag</td><td>gcgagacagc</td><td>gatgttaagc</td><td>1260</td>

<td>gaaccccaaa</td><td>aataacgtcc</td><td>cagttcggat</td><td>tgcagcctgc</td><td>aactcggctg</td><td>catgaagctg</td><td>1320</td>

<td>gaatcgctag</td><td>taatcgcgga</td><td>tcagaatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>tcttgtacac</td><td>1380</td>

<td>accgcccgtc</td><td>acaccatggg</td><td>agtcgggaac</td><td>gcccgaagcc</td><td>ggtgaccgaa</td><td>cccgaaaggg</td><td>1440</td>

<td>gaggagccgt</td><td>cgaaggcggg</td><td>cctggtgact</td><td>ggggtgaagt</td><td>cgtaacaagg</td><td>tagccgt</td><td>1497</td>

<210> 51 <211> 1475 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1475) <223> Sekwencja genu kodującego 16S rRNA 31 szczepu Clostridium <400> 51

<td>gagtttgatc</td><td>ctggctcagg</td><td>ataaacgctg</td><td>gcggcgcaca</td><td>taagacatgc</td><td>aagtcgaacg</td><td>60</td>

<td>aacttaatac</td><td>cttgcttgca</td><td>aggtaagcgg</td><td>ttagtggcgg</td><td>actggtgagt</td><td>aacacgtaag</td><td>120</td>

<td>aaatctgcct</td><td>atcagagggg</td><td>aataacagtg</td><td>agaaatcact</td><td>gctaataccg</td><td>catatgccat</td><td>180</td>

<td>agttatcgca</td><td>tgataatagt</td><td>gggaaagaag</td><td>caattcgctg</td><td>atagatgagc</td><td>ttgcggctga</td><td>240</td>

<td>ttagctagtt</td><td>ggtggggtaa</td><td>cggcctacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>cctgagaggg</td><td>. 300</td>

<td>tgaacggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccaaactcct</td><td>acgggaggca</td><td>gcagtgggga</td><td>360</td>

<td>atattgcaca</td><td>atgggggaaa</td><td>ccctgatgca</td><td>gcgacgccgc</td><td>gtgagtgaag</td><td>aagtatttcg</td><td>420</td>

<td>gtatgtaaag</td><td>ctctatcagc</td><td>agggaagaaa</td><td>atgacggtac</td><td>ctgactaaga</td><td>aagccccggc</td><td>480</td>

<td>taactacgtg</td><td>ccagcagccg</td><td>cggtaatacg</td><td>tagggggcaa</td><td>gcgttatccg</td><td>gatttactgg</td><td>540</td>

<td>tgtaaaggga</td><td>gcgtagacgg</td><td>cagcgcaagt</td><td>ctgagtgaaa</td><td>tcccatggct</td><td>tacccatgaa</td><td>600</td>

<td>actgctttgg</td><td>aaactgtgca</td><td>gctggagtgc</td><td>aggagaggta</td><td>agcggaatcc</td><td>tagtgtagcg</td><td>660</td>

<td>gttgaaatgc</td><td>gtagattatc</td><td>agaaggaaca</td><td>ccggtggccg</td><td>aggcggcctg</td><td>ctgggctttt</td><td>720</td>

<td>actgacgctg</td><td>aggctcgaag</td><td>cgtgggtagc</td><td>aaacaggatt</td><td>agataccctg</td><td>gtagtccacg</td><td>780</td>

<td>cggtaaacga</td><td>tgattactag</td><td>gtgtgggtgg</td><td>actgacccca</td><td>tccgtgccgg</td><td>agttaacaca</td><td>840</td>

<td>ataagtaatc</td><td>cacctgggga</td><td>gtacggccgc</td><td>aaggttgaaa</td><td>ctcaaaggaa</td><td>ttgacggggg</td><td>900</td>

<td>cccgcacaag</td><td>cagtggagta</td><td>tgtggtttaa</td><td>ttcgacgcaa</td><td>cgcgaagaac</td><td>cttaccaggt</td><td>960</td>

<td>cttgacatcg</td><td>agtgacggac</td><td>atagagatat</td><td>gtctttcctt</td><td>cgggacacga</td><td>agacaggtgg</td><td>1020</td>

<td>tgcatggttg</td><td>tcgtcagctc</td><td>gtgtcgtgag</td><td>atgttgggtt</td><td>aagtcccgca</td><td>acgagcgcaa</td><td>1080</td>

<td>cccttaccat</td><td>tagttgctac</td><td>gcaagagcac</td><td>tctaatggga</td><td>ctgccgttga</td><td>caaaacggag</td><td>1140</td>

<td>gaaggtgggg</td><td>atgacgtcaa</td><td>atcatcatgc</td><td>cccttatgac</td><td>ctgggcgaca</td><td>cacgtactac</td><td>1200</td>

<td>aatggcggtc</td><td>aacagaggga</td><td>ggcaaagccg</td><td>cgaggcagag</td><td>caaaccccta</td><td>aaagccgtct</td><td>1260</td>

<td>cagttcggat</td><td>tgcaggctgc</td><td>aactcgcctg</td><td>catgaagtcg</td><td>gaattgctag</td><td>taatcgcgga</td><td>1320</td>

<td>tcagcatgcc</td><td>gcggtgaata</td><td>cgttcccggg</td><td>ccttgtacac</td><td>accgcccgtc</td><td>acaccatgag</td><td>1380</td>

<td>agccggtaac</td><td>acccgaagtc</td><td>aatagtctaa</td><td>ccgcaaggag</td><td>gacattgccg</td><td>aaggtgggat</td><td>1440</td>

<td>tggtaattgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td></td><td></td><td>1475</td>

<210> 52 <211> 1491 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1491) <223> Sekwencja genu kodującego 16S rRNA szczepu Clostridium 32 <400> 52

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gagaatcagt</td><td>ggattgagga</td><td>ttcgtccaaa</td><td>tgaaactgag</td><td>gaaagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtga</td><td>gcaatctgcc</td><td>ttggagtggg</td><td>gaataacggc</td><td>tggaaacagc</td><td>cgctaatacc</td><td>180</td>

<td>gcatgataca</td><td>gttgggaggc</td><td>atctctctga</td><td>ctgtcaaaga</td><td>tttatcgctc</td><td>tgagatgagc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggcggggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atgggcgcaa</td><td>gcctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttctg</td><td>ggggacgaac</td><td>aaatgacggt</td><td>accccaggaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttatcc</td><td>540</td>

<td>ggatttattg</td><td>ggtgtaaagg</td><td>gcgtgtaggc</td><td>gggaatgcaa</td><td>gtcagatgtg</td><td>aaaactatgg</td><td>600</td>

<td>gctcaaccca</td><td>tagcctgcat</td><td>ttgaaactgt</td><td>atttcttgag</td><td>tgctggagag</td><td>gcaatcggaa</td><td>660</td>

<td>ttccgtgtgt</td><td>agcggtgaaa</td><td>tgcgtagata</td><td>tacggaggaa</td><td>caccagtggc</td><td>gaagcggatt</td><td>720</td>

<td>gctggacagt</td><td>aactgacgct</td><td>gaggcgcgaa</td><td>agcgtgggga</td><td>gcaaacagga</td><td>ttagataccc</td><td>780</td>

<td>tggtagtcca</td><td>cgccgtaacg</td><td>atggatacta</td><td>gtgtgggggg</td><td>actgaccccc</td><td>tccgtgccgc</td><td>840</td>

<td>agctaacgca</td><td>ataagtatcc</td><td>ccacctgggg</td><td>agtacgatcg</td><td>caaggttgaa</td><td>actcaaagga</td><td>900</td>

<td>attgacgggg</td><td>gcccgcacaa</td><td>gcggtggagt</td><td>atgtggttta</td><td>attcgaagca</td><td>acgcgaagaa</td><td>960</td>

<td>ccttaccagg</td><td>gcttgacatc</td><td>ctgctaacga</td><td>accagagatg</td><td>gattaggtgc</td><td>ccttcgggga</td><td>1020</td>

<td>aagcagagac</td><td>aggtggtgca</td><td>tggttgtcgt</td><td>cagctcgtgt</td><td>cgtgagatgt</td><td>tgggttaagt</td><td>1080</td>

<td>cccgcaacga</td><td>gcgcaacccc</td><td>tattgttagt</td><td>tgctacgcaa</td><td>gagcactcta</td><td>gcgagactgc</td><td>1140</td>

<td>cgttgacaaa</td><td>acggaggaag</td><td>gtggggacga</td><td>cgtcaaatca</td><td>tcatgcccct</td><td>tacgtcctgg</td><td>1200</td>

<td>gccacacacg</td><td>tactacaatg</td><td>gcggttaaca</td><td>aagagaggca</td><td>agaccgcgag</td><td>gtggagcaaa</td><td>1260</td>

<td>tctcaaaaag</td><td>ccgtcccagt</td><td>tcggatcgca</td><td>ggctgcaacc</td><td>cgcctgcgtg</td><td>aagttggaat</td><td>1320</td>

<td>cgctagtaat</td><td>cgcggatcag</td><td>catgccgcgg</td><td>tgaatacgtt</td><td>cccgggcctt</td><td>gtacacaccg</td><td>1380</td>

<td>cccgtcacac</td><td>catgagagtc</td><td>gggaacaccc</td><td>gaagtccgta</td><td>gcctaaccgc</td><td>aaggggggcg</td><td>1440</td>

<td>cggccgaagg</td><td>tgggttcgat</td><td>aattggggtg</td><td>aagtcgtaac</td><td>aaggtagccg</td><td>t</td><td>1491</td>

<210> 53 <211> 1495 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1)..(1495) <223> Sekwencja genu kodującego 16S rRNA 33 szczepu Clostridium <400> 53

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggtgtacgg</td><td>ggaggaaggc</td><td>ttcggccgga</td><td>aaacctgtgc</td><td>atgagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtgg</td><td>gcaacctggc</td><td>ctgtacaggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>180</td>

<td>gcataacggg</td><td>ggaagccgca</td><td>tggcttttcc</td><td>ctgaaaactc</td><td>cggtggtaca</td><td>ggatgggccc</td><td>240</td>

<td>gcgtctgatt</td><td>agccagttgg</td><td>cagggtaacg</td><td>gcctaccaaa</td><td>gcgacgatca</td><td>gtagccggcc</td><td>300</td>

<td>tgagagggcg</td><td>gacggccaca</td><td>ctgggactga</td><td>gacacggccc</td><td>agactcctac</td><td>gggaggcagc</td><td>360</td>

<td>agtgggggat</td><td>attgcacaat</td><td>ggggggaacc</td><td>ctgatgcagc</td><td>gacgccgcgt</td><td>gggtgaagaa</td><td>420</td>

<td>gcgcctcggc</td><td>gcgtaaagcc</td><td>ctgtcagcag</td><td>ggaagaaaat</td><td>gacggtacct</td><td>gaagaagaag</td><td>480</td>

<td>ccccggctaa</td><td>ctacgtgcca</td><td>gcagccgcgg</td><td>taatacgtag</td><td>ggggcaagcg</td><td>ttatccggat</td><td>540</td>

<td>ttactgggtg</td><td>taaagggggc</td><td>gcagacggcg</td><td>atgcaagcca</td><td>ggagtgaaag</td><td>cccggggccc</td><td>600</td>

<td>aaccccggga</td><td>ctgctcttgg</td><td>aactgcgtgg</td><td>ctggagtgca</td><td>ggaggggcag</td><td>gcggaattcc</td><td>660</td>

<td>tggtgtagcg</td><td>gtgaaatgcg</td><td>tagatatcag</td><td>aggaacaccg</td><td>gtggcgaaag</td><td>cggcctgctg</td><td>720</td>

<td>gactgcaact</td><td>gacgttgagg</td><td>cccgaaagcg</td><td>gtgggagcaa</td><td>acaggattag</td><td>ataccctggt</td><td>780</td>

<td>agtccacgcc</td><td>gtaaacgatg</td><td>attactaggt</td><td>gtcggggagc</td><td>agagactgcc</td><td>cggtgccgca</td><td>840</td>

<td>gcccaacgca</td><td>ttaagtatcc</td><td>acctggggag</td><td>tacgttcgca</td><td>agaatgaaac</td><td>tcaaaggaat</td><td>900</td>

<td>tgacggggac</td><td>ccgcacaagc</td><td>ggtggagcat</td><td>gtggtttaat</td><td>tcgaagcaac</td><td>gcgaagaacc</td><td>960</td>

<td>ttaccaggcc</td><td>ttgacatccc</td><td>cctggatggc</td><td>ccgtaacggg</td><td>gccagccctt</td><td>tttgggcagg</td><td>1020</td>

<td>ggagacaggt</td><td>ggtgcatggt</td><td>tgtcgtcagc</td><td>tcgtgtcgtg</td><td>agatgttggg</td><td>ttaagtcccg</td><td>1080</td>

<td>caacgagcgc</td><td>aacccctgcc</td><td>cgcagtagcc</td><td>agcattttag</td><td>atggggactc</td><td>tgcggggact</td><td>1140</td>

<td>gccggggaca</td><td>acccggagga</td><td>aggcggggat</td><td>gacgtcaaat</td><td>catcatgccc</td><td>cttatggcct</td><td>1200</td>

<td>gggctacaca</td><td>cgtgctacaa</td><td>tggcgccgac</td><td>agagggagga</td><td>gaagcggcga</td><td>cgcggagcga</td><td>1260</td>

<td>accccaaaaa</td><td>cggcgtccca</td><td>gttcggattg</td><td>tagtctgcaa</td><td>cccgactaca</td><td>tgaagccgga</td><td>1320</td>

<td>atcgctagta</td><td>atcgcggatc</td><td>agaatgccgc</td><td>ggtgaatacg</td><td>ttcccgggtc</td><td>ttgtacacac</td><td>1380</td>

<td>cgcccgtcac</td><td>accatgggag</td><td>ccgggaatgc</td><td>ccgaagtctg</td><td>tgaccgaacc</td><td>cgtaagggga</td><td>1440</td>

<td>ggggcagccg</td><td>aaggcaggcc</td><td>cggtgactgg</td><td>ggtgaagtcg</td><td>taacaaggta</td><td>gccgt</td><td>1495</td>

<210> 54 <211> 1493 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1493) <223> Sekwencja genu kodującego 16S rRNA 34 szczepu Clostridium <400> 54

<td>agagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatgggaat</td><td>gcttagtgtc</td><td>ggactggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>cttccagagg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcatgatgc</td><td>gttggagccg</td><td>catgactccg</td><td>acgtcaaaga</td><td>tttatcgctg</td><td>gaagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>tttgttagtt</td><td>ggtgaggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atggacgcaa</td><td>gtctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>ggggaagagc</td><td>agaagacggt</td><td>accccttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttgtcc</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gcgtgcagcc</td><td>ggagagacaa</td><td>gtcagatgtg</td><td>aaatccacgg</td><td>600</td>

<td>gctcaacccg</td><td>tgaactgcat</td><td>ttgaaactgt</td><td>ttcccttgag</td><td>tgtcggagag</td><td>gtaatcggaa</td><td>660</td>

<td>ttccttgtgt</td><td>agcggtgaaa</td><td>tgcgtagata</td><td>taaggaagaa</td><td>caccagtggc</td><td>gaaggcggat</td><td>720</td>

<td>tactggacga</td><td>taactgacgg</td><td>tgaggcgcga</td><td>aagcgtgggg</td><td>agcaaacagg</td><td>attagatacc</td><td>780</td>

<td>ctggtagtcc</td><td>acgctgtaaa</td><td>cgatcgatac</td><td>taggtgtgcg</td><td>gggactgacc</td><td>ccctgcgtgc</td><td>840</td>

<td>cggagttaac</td><td>acaataagta</td><td>tcgcacctgg</td><td>ggagtacgat</td><td>cgcaaggttg</td><td>aaactcaaag</td><td>900</td>

<td>gaattgacgg</td><td>gggcccgcac</td><td>aagcggtgga</td><td>ttatgtggtt</td><td>taattcgaag</td><td>caacgcgaag</td><td>960</td>

<td>aaccttacca</td><td>gggcttgaca</td><td>tcctgctaac</td><td>gaagtagaga</td><td>tacattaggt</td><td>gcccttcggg</td><td>1020</td>

<td>gaaagcagag</td><td>acaggtggtg</td><td>catggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>gttgggttaa</td><td>1080</td>

<td>gtcccgcaac</td><td>gagcgcaacc</td><td>cctattgtta</td><td>gttgctacgc</td><td>aagagcactc</td><td>tagcgagact</td><td>1140</td>

<td>gccgttgaca</td><td>aaacggagga</td><td>aggcggggac</td><td>gacgtcaaat</td><td>catcatgccc</td><td>cttatgtcct</td><td>1200</td>

<td>gggctacaca</td><td>cgtaatacaa</td><td>tggcggttaa</td><td>caaagggatg</td><td>caaagccgcg</td><td>aggcagagcg</td><td>1260</td>

<td>aaccccaaaa</td><td>agccgtccca</td><td>gttcggatcg</td><td>caggctgcaa</td><td>cccgcctgcg</td><td>tgaagtcgga</td><td>1320</td>

<td>atcgctagta</td><td>atcgcggatc</td><td>agcatgccgc</td><td>ggtgaatacg</td><td>ttcccgggcc</td><td>ttgtacacac</td><td>1380</td>

<td>cgcccgtcac</td><td>accatgagag</td><td>tcgggaacac</td><td>ccgaagtccg</td><td>tagcctaacc</td><td>gcaaggaggg</td><td>1440</td>

<td>cgcggccgaa</td><td>ggtgggttcg</td><td>ataattgggg</td><td>tgaagtcgta</td><td>acaaggtagc</td><td>cgt</td><td>1493</td>

<210> 55 <211> 1498 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1)..(1498) <223> Sekwencja genu kodującego 16S rRNA 35 szczepu Clostridium <400> 55

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggtgtacag</td><td>aagggaagat</td><td>tacggtcgga</td><td>aggtctgtgc</td><td>atgagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtgg</td><td>gcaacctggc</td><td>ctgtacaggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>180</td>

<td>gcataacggg</td><td>ggaagccgca</td><td>tggcttttcc</td><td>ctgaaaactc</td><td>cggtggtaca</td><td>ggatgggccc</td><td>240</td>

<td>gcgtctgatt</td><td>attttttttg</td><td>tcagggtaac</td><td>ggcctaccaa</td><td>agcgacgatc</td><td>agtagccggc</td><td>300</td>

<td>ctgagagggc</td><td>ggacggccac</td><td>actgggactg</td><td>agacacggcc</td><td>cagactccta</td><td>cgggaggcag</td><td>360</td>

<td>cagtggggga</td><td>tattgcacaa</td><td>tggggggaac</td><td>cctgatgcag</td><td>cgacgccgcg</td><td>tgggtgaaga</td><td>420</td>

<td>agcgcctcgg</td><td>cgcgtaaagc</td><td>cctgtcagca</td><td>gggaagaaaa</td><td>tgacggtacc</td><td>tgaagaagaa</td><td>480</td>

<td>gccccggcta</td><td>actacgtgcc</td><td>agcagccgcg</td><td>gtaatacgta</td><td>aggggcaagc</td><td>gttatccgga</td><td>540</td>

<td>tttactgggt</td><td>gtaaaggggg</td><td>cgcagacggc</td><td>gatgcaagcc</td><td>aggagtgaaa</td><td>gcccggggcc</td><td>600</td>

<td>caaccccggg</td><td>actgctcttg</td><td>ggaactgcgg</td><td>tggctggagt</td><td>gcaggagggg</td><td>caggccggaa</td><td>660</td>

<td>ttcctggtgt</td><td>agcggtgaaa</td><td>tgcgtagata</td><td>tcaggaggaa</td><td>caccggtggc</td><td>gaaggcggcc</td><td>720</td>

<td>tgctggactg</td><td>caactgacgt</td><td>tgaggcccga</td><td>aagcgtgggg</td><td>agcaaacagg</td><td>attagatacc</td><td>780</td>

<td>ctggtagtca</td><td>cgccgtaaac</td><td>gatgattact</td><td>aggtgtcggg</td><td>gagcagagac</td><td>tgcccggtgc</td><td>840</td>

<td>cgcagccaac</td><td>gcattaagta</td><td>atccacctgg</td><td>ggagtacgtt</td><td>cgcaagaatg</td><td>aaactcaaag</td><td>900</td>

<td>gaattgacgg</td><td>ggacccgcac</td><td>aagcggtgga</td><td>gcatgtggtt</td><td>taattcgaag</td><td>caacgcgaag</td><td>960</td>

<td>aaccttacca</td><td>ggccttgaca</td><td>tccccctgga</td><td>tggcccgtaa</td><td>cggggtcagc</td><td>ctttcggggc</td><td>1020</td>

<td>aggggagaca</td><td>ggtggtgcat</td><td>ggttgtcgtc</td><td>agctcgtgtc</td><td>gtgagatgtt</td><td>gggttaagtc</td><td>1080</td>

<td>ccgcaacgag</td><td>cgcaacccct</td><td>gcccgcagta</td><td>gccagcattt</td><td>tagatgggga</td><td>ctctgcgggg</td><td>1140</td>

<td>actgccgggg</td><td>acaacccgga</td><td>ggaaggcggg</td><td>gatgacgtca</td><td>aatcatcatg</td><td>ccccttatgg</td><td>1200</td>

<td>cctgggctac</td><td>acacgtgcta</td><td>caatggcgcc</td><td>gacagaggga</td><td>ggcgaagcgg</td><td>cgacgcggag</td><td>1260</td>

<td>cgaaccccaa</td><td>aaacggcgtc</td><td>ccagttcgga</td><td>ttgtagtctg</td><td>caacccgact</td><td>acatgaagcc</td><td>1320</td>

<td>ggaatcgcta</td><td>gtaatcgcgg</td><td>atcagaatgc</td><td>cgcggtgaat</td><td>acgttcccgg</td><td>gtcttgtaca</td><td>1380</td>

<td>caccgcccgt</td><td>cacaccatgg</td><td>gagccgggaa</td><td>tgcccgaagt</td><td>ctgtgaccga</td><td>acccgtaagg</td><td>1440</td>

<td>ggaggggcag</td><td>ccgaaggcag</td><td>gcccggtgac</td><td>tggggtgaag</td><td>tcgtaacaag</td><td>gtagccgt</td><td>1498</td>

<210> 56 <211> 1491 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1491) <223> Sekwencja genu kodującego 16S rRNA 36 szczepu Clostridium <400> 56

<td>agagtttgat</td><td>catggctcag</td><td>gacgaacgct</td><td>ggcggcaagc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcgccta</td><td>tgaaggagat</td><td>ttcggtcaac</td><td>ggaataggct</td><td>gcttagtggc</td><td>tgacgggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>ctttcagagg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcataacac</td><td>ataggtgtcg</td><td>catggcattt</td><td>atgtcaaaga</td><td>tttatcgctg</td><td>aaagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggtgaggtaa</td><td>cggctcacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tagccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atggacgcaa</td><td>gtctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>agggaagagc</td><td>agaagacggt</td><td>acctcttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtagtggcaa</td><td>gcgttgtccg</td><td>540</td>

<td>gatttactgg</td><td>gtgtaaaggg</td><td>cgtgtagccg</td><td>ggctgacagt</td><td>cagatgtgaa</td><td>attccggggc</td><td>600</td>

<td>tcaaccccgg</td><td>acctgcattt</td><td>gaaactgttg</td><td>gtcttgagta</td><td>tcggagaggc</td><td>aggcggaatt</td><td>660</td>

<td>cctagtgtag</td><td>cggtgaaatg</td><td>cgtagatatt</td><td>aggaggaaca</td><td>ccagtggcga</td><td>aggcggcctg</td><td>720</td>

<td>ctggacgaca</td><td>actgacggtg</td><td>aggcgcgaaa</td><td>gcgtggggag</td><td>caaacaggat</td><td>tagataccct</td><td>780</td>

<td>ggtagtccac</td><td>gctgtaaacg</td><td>atggatacta</td><td>ggtgtgcggg</td><td>gactgacccc</td><td>ctgcgtgccg</td><td>840</td>

<td>cagttaacac</td><td>aataagtatc</td><td>ccacctgggg</td><td>agtacgatcg</td><td>caaggttgaa</td><td>actcaaagga</td><td>900</td>

<td>attgacgggg</td><td>gcccgcacaa</td><td>gcggtggatt</td><td>atgtggttta</td><td>attcgatgca</td><td>acgcgaagaa</td><td>960</td>

<td>ccttaccagg</td><td>gcttgacatc</td><td>ctgctaacga</td><td>ggtagagata</td><td>cgtcaggtgc</td><td>ccttcgggga</td><td>1020</td>

<td>aagcagagac</td><td>aggtggtgca</td><td>tggttgtcgt</td><td>cagctcgtgt</td><td>cgtgagatgt</td><td>tgggttaagt</td><td>1080</td>

<td>cccgcaacga</td><td>gcgcaaccct</td><td>tattgttagt</td><td>tgctacgcaa</td><td>gagcactcta</td><td>gcgagactgc</td><td>1140</td>

<td>cgttgacaaa</td><td>acggaggaag</td><td>gtggggacga</td><td>cgtcaaatca</td><td>tcatgcccct</td><td>tatgtcctgg</td><td>1200</td>

<td>gctacacacg</td><td>taatacaatg</td><td>gcggtaaaca</td><td>gagggatgca</td><td>atactgcgaa</td><td>gtggagcgaa</td><td>1260</td>

<td>cccctaaaag</td><td>ccgtcccagt</td><td>tcagattgca</td><td>gtctgcaact</td><td>cgactgcatg</td><td>aagtcggaat</td><td>1320</td>

<td>cgctagtaat</td><td>cgcggatcag</td><td>catgccgcgg</td><td>tgaatacgtt</td><td>cccgggcctt</td><td>gtacacaccg</td><td>1380</td>

<td>cccgtcacac</td><td>catgagagtc</td><td>gggaacaccc</td><td>gaagtccgta</td><td>gcctaaccgc</td><td>aaggagggcg</td><td>1440</td>

<td>cggccgaagg</td><td>tgggttcgat</td><td>aattggggtg</td><td>aagtcgtaac</td><td>aaggtagccg</td><td>t</td><td>1491</td>

<210> 57 <211> 1493 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1)..(1493) <223> Sekwencja genu kodującego 16S rRNA 37 szczepu Clostridium <400> 57

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>gggtgtacgg</td><td>ggaggaaggc</td><td>ttcggccgga</td><td>aaacctgtgc</td><td>atgagtggcg</td><td>gacgggtgag</td><td>120</td>

<td>taacgcgtgg</td><td>gcaacctggc</td><td>ctgtacaggg</td><td>ggataacact</td><td>tagaaatagg</td><td>tgctaatacc</td><td>180</td>

<td>gcataacggg</td><td>ggaagccgca</td><td>tggcttttcc</td><td>ctgaaaactc</td><td>cggtggtaca</td><td>ggatgggccc</td><td>240</td>

<td>gcgtctgatt</td><td>agccagttgg</td><td>cagggtaacg</td><td>gcctaccaaa</td><td>gcgacgatca</td><td>gtagccggcc</td><td>300</td>

<td>tgagagggcg</td><td>gacggccaca</td><td>ctgggactga</td><td>gacacggccc</td><td>agactcctac</td><td>gggaggcagc</td><td>360</td>

<td>agtgggggat</td><td>attgcacaat</td><td>ggggggaaac</td><td>cctgatgcag</td><td>cgacgccgcg</td><td>tgagtgaaga</td><td>420</td>

<td>agtatttcgg</td><td>tatgtaaagc</td><td>tctatcagca</td><td>gggaagaaaa</td><td>tgacggtacc</td><td>tgactaagaa</td><td>480</td>

<td>gccccggcta</td><td>actacgtgcc</td><td>agcagccgcg</td><td>gtaatacgta</td><td>gggggcaagc</td><td>gttatccgga</td><td>540</td>

<td>tttactgggt</td><td>gtaaagggag</td><td>cgtagacggc</td><td>agcgcaagtc</td><td>tgaagtgaaa</td><td>tcccatggct</td><td>600</td>

<td>taaccatgga</td><td>actgctttgg</td><td>aaactgtgca</td><td>gctggagtgc</td><td>aggagaggta</td><td>agcggaattc</td><td>660</td>

<td>ctagtgtagc</td><td>ggtgaaatgc</td><td>gtagatatta</td><td>ggaggaacac</td><td>cagtggcgaa</td><td>ggcggcttac</td><td>720</td>

<td>tggactgtac</td><td>tgacgttgag</td><td>gctcgaaagc</td><td>gtggggagca</td><td>aacaggatta</td><td>gataccctgg</td><td>780</td>

<td>tagtccacgc</td><td>cgtaaacgat</td><td>gattactagg</td><td>tgttggggga</td><td>ccaaggtctt</td><td>cggtgccggc</td><td>840</td>

<td>gcaaacgcat</td><td>taagtaatcc</td><td>acctggggag</td><td>tacgttcgca</td><td>agaatgaaac</td><td>tcaaaggaat</td><td>900</td>

<td>tgacggggac</td><td>ccgcacaagc</td><td>ggtggagcat</td><td>gtggtttaat</td><td>tcgaagcaac</td><td>gcgaagaacc</td><td>960</td>

<td>ttacctggtc</td><td>ttgacatccc</td><td>gatgacgagt</td><td>gagcaaagtc</td><td>actttccctt</td><td>cggggcattg</td><td>1020</td>

<td>gagacaggtg</td><td>gtgcatggtt</td><td>gtcgtcagct</td><td>cgtgtcgtga</td><td>gatgttgggt</td><td>taagtcccgc</td><td>1080</td>

<td>aacgagcgca</td><td>acccctattt</td><td>ccagtagcca</td><td>gcaggtagag</td><td>ctgggcactc</td><td>tggagagact</td><td>1140</td>

<td>gcccgggata</td><td>accgggagga</td><td>aggcggggat</td><td>gacgtcaaat</td><td>catcatgccc</td><td>cttatgatca</td><td>1200</td>

<td>gggctacaca</td><td>cgtgctacaa</td><td>tggcgtaaac</td><td>aaagggaagc</td><td>gagacggtga</td><td>cgttgagcaa</td><td>1260</td>

<td>atcccaaaaa</td><td>taacgtccca</td><td>gttcggattg</td><td>tagtctgcaa</td><td>ctcgactaca</td><td>tgaagctgga</td><td>1320</td>

<td>atcgctagta</td><td>atcgcgaatc</td><td>agaatgtcgc</td><td>ggtgaatacg</td><td>ttcccgggtc</td><td>ttgtacacac</td><td>1380</td>

<td>cgcccgtcac</td><td>accatgggag</td><td>tcggaaatgc</td><td>ccgaagtcag</td><td>tgacctaacc</td><td>gaaaggaagg</td><td>1440</td>

<td>agctgccgaa</td><td>ggtggagccg</td><td>gtaactgggg</td><td>tgaagtcgta</td><td>acaaggtagc</td><td>cgt</td><td>1493</td>

<210> 58 <211> 1493 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1493) <223> Sekwencja genu kodującego 16S rRNA 38 szczepu Clostridium <400> 58

<td>aaagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatgggaat</td><td>gcttagtggc.</td><td>ggactggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>cttccagagg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcatgatgc</td><td>gttggagccg</td><td>catgactccg</td><td>acgtcaaaga</td><td>.tttatcgctg</td><td>gaagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggtgaggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atggacgcaa</td><td>gtctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>ggggaagagc</td><td>agaagacggt</td><td>accccttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtagtggcaa</td><td>gcgttgtccg</td><td>540</td>

<td>gatttactgg</td><td>gtgtaaaggg</td><td>cgtgcagccg</td><td>gagagacaag</td><td>tcagatgtga</td><td>aatccacggg</td><td>600</td>

<td>ctcaacccgt</td><td>gaactgcatt</td><td>tgaaactgtt</td><td>tcccttgagt</td><td>gtcggagagg</td><td>taatcggaat</td><td>660</td>

<td>tccttgtgta</td><td>gcggtgaaat</td><td>gcgtagatat</td><td>aaggaagaac</td><td>accagtggcg</td><td>aaggcggatt</td><td>720</td>

<td>actggacgat</td><td>aaactgacgg</td><td>tgaggcgcga</td><td>aagcgtgggg</td><td>agcaaacagg</td><td>attagatacc</td><td>780</td>

<td>ctggtagtcc</td><td>acgctgtaaa</td><td>cgatcgatac</td><td>taggtgtgcg</td><td>gggactgacc</td><td>ccctgcgtgc</td><td>840</td>

<td>cggagttaac</td><td>acaataagta</td><td>tcgcacctgg</td><td>ggagtacgat</td><td>cgcaaggttg</td><td>aaactcaaag</td><td>900</td>

<td>gaattgacgg</td><td>gggcccgcac</td><td>aagcggtgga</td><td>ttatgtggtt</td><td>taattcgaag</td><td>caacgcgaag</td><td>960</td>

<td>aaccttacca</td><td>gggcttgaca</td><td>tcctgctaac</td><td>gaagtagaga</td><td>tacattaggt</td><td>gcccttcggg</td><td>1020</td>

<td>gaaagtagag</td><td>acaggtggtg</td><td>catggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>gttgggttaa</td><td>1080</td>

<td>gtcccgcaac</td><td>gagcgcaacc</td><td>cctattgtta</td><td>gttgctacgc</td><td>aagagcactc</td><td>tagcgagact</td><td>1140</td>

<td>gccgttgaca</td><td>aaacggagga</td><td>aggcggggac</td><td>gacgtcaaat</td><td>catcatgccc</td><td>cttatgtcct</td><td>1200</td>

<td>gggctacaca</td><td>cgtaatacaa</td><td>tggcggttaa</td><td>caaagggatg</td><td>caaagccgcg</td><td>aggcagagcg</td><td>1260</td>

<td>aaccccaaaa</td><td>agccgtccca</td><td>gttcggatcg</td><td>caggctgcaa</td><td>cccgcctgcg</td><td>tgaagtcgga</td><td>1320</td>

<td>atcgctagta</td><td>atcgcggatc</td><td>agcatgccgc</td><td>ggtgaatacg</td><td>ttcccgggcc</td><td>ttgtacacac</td><td>1380</td>

<td>cgcccgtcac</td><td>accatgagag</td><td>tcgggaacac</td><td>ccgaagtccg</td><td>tagcctaacc</td><td>gcaaggaggg</td><td>1440</td>

<td>cgcggccgaa</td><td>ggtgggttcg</td><td>ataattgggg</td><td>tgaagtcgta</td><td>acaaggtagc</td><td>cgt</td><td>1493</td>

<210> 59 <211> 1511 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1511) <223> Sekwencja genu kodującego 16S rRNA 39 szczepu Clostridium <400> 59

<td>agagtttgat</td><td>cctggctcag</td><td>gacgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcacccc</td><td>tgaaggagtt</td><td>ttcggacaac</td><td>ggatgggaat</td><td>gcttagtggc</td><td>ggactggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>cttccagagg</td><td>gggacaacag</td><td>ttggaaacga</td><td>ctgctaatac</td><td>180</td>

<td>cgcatgatgc</td><td>gttggagccg</td><td>catgactccg</td><td>acgtcaaaga</td><td>tttatcgctg</td><td>gaagatggcc</td><td>240</td>

<td>tcgcgtctga</td><td>ttagctagtt</td><td>ggtgaggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>cagtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tggccggcca</td><td>cattgggact</td><td>gagatacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgggca</td><td>atggacgcaa</td><td>gtctgaccca</td><td>gcaacgccgc</td><td>gtgaaggaag</td><td>420</td>

<td>aaggctttcg</td><td>ggttgtaaac</td><td>ttcttttaag</td><td>ggggaagagc</td><td>agaagacggt</td><td>accccttgaa</td><td>480</td>

<td>taagccacgg</td><td>ctaactacgt</td><td>gccagcagcc</td><td>gcggtaatac</td><td>gtaggtggca</td><td>agcgttgtcc</td><td>540</td>

<td>ggatttactg</td><td>ggtgtaaagg</td><td>gcgtgcagcc</td><td>ggagagacaa</td><td>gtcagatgtg</td><td>aaatccacgg</td><td>600</td>

<td>gctcaacccg</td><td>tgaactgcat</td><td>ttgaaactgt</td><td>ttcccttgag</td><td>tgtcggagag</td><td>gtaatcggaa</td><td>660</td>

<td>ttccttgtgt</td><td>agcggtgaaa</td><td>tgcgtagata</td><td>taaggaagac</td><td>accagtggcg</td><td>aagcggatta</td><td>720</td>

<td>ctggacgata</td><td>actgacggtg</td><td>aggcgcgaaa</td><td>gcgtggggag</td><td>caaacaggat</td><td>tagatacctg</td><td>780</td>

<td>ggtagtcaac</td><td>gctgtaaacg</td><td>atcgatacta</td><td>ggtggtgcgg</td><td>gggacttgac</td><td>cccctgccgt</td><td>840</td>

<td>tgccggagtt</td><td>aacaccaata</td><td>aagtattcgg</td><td>caccctgggg</td><td>agtacgatcg</td><td>caaaggttga</td><td>900</td>

<td>aaactcaaaa</td><td>gaaatggacg</td><td>gggggccccg</td><td>ccccaagcgg</td><td>gtgggattat</td><td>gttggtttat</td><td>960</td>

<td>ttcgaaagca</td><td>acgcgaagaa</td><td>ccctaacagg</td><td>gcttgacatc</td><td>ctgctaacga</td><td>agtagagata</td><td>1020</td>

<td>cattaggtgc</td><td>ccttcgggga</td><td>aagtagagac</td><td>aggtggtgca</td><td>tggttgtcgt</td><td>cagctcgtgt</td><td>1080</td>

<td>cgtgagatgt</td><td>tgggttaagt</td><td>cccgcaacga</td><td>gcgcaacccc</td><td>tattgttagt</td><td>tgctacgcaa</td><td>1140</td>

<td>gagcactcta</td><td>gcgagactgc</td><td>cgttgacaaa</td><td>acggaggaag</td><td>gcggggacga</td><td>cgtcaaatca</td><td>1200</td>

<td>tcatgcccct</td><td>tatgtcctgg</td><td>gctacacacg</td><td>taatacaatg</td><td>gcggttaaca</td><td>aagggatgca</td><td>1260</td>

<td>aagccgcgag</td><td>gcagagcgaa</td><td>ccccaaaaag</td><td>ccgtcccagt</td><td>tcggatcgca</td><td>ggctgcaacc</td><td>1320</td>

<td>cgcctgcgtg</td><td>aagtcggaat cgctagtaat cgcggatcag catgccgcgg tgaatacgtt</td><td>1380</td>

<td>cccgggcctt</td><td>gtacacaccg cccgtcacac catgagagtc gggaacaccc gaagtccgta</td><td>1440</td>

<td>gcctaaccgc</td><td>aaggagggcg cggccgaagg tgggttcgat aattggggtg aagtcgtaac</td><td>1500</td>

<td>aaggtagccg</td><td>t</td><td>1511</td>

<210> 60 <211> 1499 <212> DNA <213> Clostridium leptum <220>

<221> rRNA <222> (1)..(1499) <223> Sekwencja genu kodującego 16S rRNA 40 szczepu Clostridium <400> 60

<td>agagtttgat</td><td>cctggctcag</td><td>gataaacgct</td><td>ggcggcatgc</td><td>ctaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagcgcctt</td><td>ggaaggagac</td><td>ttcggtcaac</td><td>ggaagaggag</td><td>gcttagtggc</td><td>ggacgggtga</td><td>120</td>

<td>gtaacgcgtg</td><td>aggaacctgc</td><td>ctcagagagg</td><td>gggataacac</td><td>accgaaaggt</td><td>gtgctaatac</td><td>180</td>

<td>cgcataacat</td><td>atgagagggg</td><td>catccctttc</td><td>atatcaaaga</td><td>tttattgctt</td><td>tgagatggcc</td><td>240</td>

<td>tcgcgtccaa</td><td>ttagctagtt</td><td>ggtgaggtaa</td><td>cggcccacca</td><td>aggcgacgat</td><td>tggtagccgg</td><td>300</td>

<td>actgagaggt</td><td>tgaacggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccagactcct</td><td>acgggaggca</td><td>360</td>

<td>gcagtgggga</td><td>atattgcaca</td><td>atggggggaa</td><td>ccctgatgca</td><td>gcaatgccgc</td><td>gtgaaggatg</td><td>420</td>

<td>aaggttttcg</td><td>gattgtaaac</td><td>ttcttttgta</td><td>cgggacgaag</td><td>aaagtgacgg</td><td>taccgtaaga</td><td>480</td>

<td>ataagccacg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtaggtggc</td><td>aagcgttatc</td><td>540</td>

<td>cggatttact</td><td>gggtgtaaag</td><td>ggcgagtagg</td><td>cgggattgca</td><td>agtcagatgt</td><td>gaaaactatg</td><td>600</td>

<td>ggctcaaccg</td><td>atagagtgca</td><td>tttgaaactg</td><td>cagttcttga</td><td>gtgatggaga</td><td>ggcaggcgga</td><td>660</td>

<td>attcccggtg</td><td>tagcggtgga</td><td>atgcgtagat</td><td>atcgggaggg</td><td>aacaccagtg</td><td>gcgaaggcgg</td><td>720</td>

<td>cctgctggac</td><td>attaactgac</td><td>gctgatgcgc</td><td>gaaagcgtgg</td><td>ggagcaaaca</td><td>ggattagata</td><td>7S0</td>

<td>ccctggtagt</td><td>cacgctgtaa</td><td>acgatgatta</td><td>ctaggtgtgg</td><td>ggggtactga</td><td>cccccttccc</td><td>840</td>

<td>gtgccggagt</td><td>taacacaata</td><td>agtaatccac</td><td>c tggggagta</td><td>cggccgcaag</td><td>gttgaaactc</td><td>900</td>

<td>aaaggaattg</td><td>acgggggccc</td><td>gcacaagcag</td><td>tggagtatgt</td><td>ggttttaatt</td><td>cgaagcaacg</td><td>960</td>

<td>cgaagaacct</td><td>taccagggct</td><td>tgacatgggg</td><td>atgaccgctt</td><td>tagagataga</td><td>gctttctctt</td><td>1020</td>

<td>cggagacatc</td><td>ccacacaggt</td><td>ggtgcatggt</td><td>tgtcgtcagc</td><td>tcgtgtcgtg</td><td>agatgttggg</td><td>1080</td>

<td>ttaagtcccg</td><td>caacgagcgc</td><td>aacccttatt</td><td>gttagttgct</td><td>acgcaagagc</td><td>actctagcga</td><td>1140</td>

<td>gactgccgtt</td><td>gacaaaacgg</td><td>aggaaggtgg</td><td>ggacgacgtc</td><td>aaatcatcat</td><td>gccctttatg</td><td>1200</td>

<td>tcctgggcta</td><td>cacacgtact</td><td>acaatggcgg</td><td>acatacagag</td><td>ggaagcaaga</td><td>cagcgatgtg</td><td>1260</td>

<td>gagcaaatcc</td><td>ctaaaagccg</td><td>tctcagttca</td><td>gattgcaggc</td><td>tgcaacccgc</td><td>ctgcatgaag</td><td>1320</td>

<td>tcggaattgc</td><td>tagtaatcgc</td><td>ggatcagcat</td><td>gccgcggtga</td><td>atacgttccc</td><td>gggccttgta</td><td>1380</td>

cacaccgccc gtcacaccat gagagtcgga aacacccgaa gcctgtagcc caaccgcaag 1440 gggggcgcag tcgaaggtgg gtctgataat tggggtgaag tcgtaacaaa ggtagccgt 1499 <210> 61 <211> 1512 <212> DNA <213> Clostridium coccoides <220>

<221> rRNA <222> (1)..(1512) <223> Sekwencja genu kodującego 16S rRNA 41 szczepu Clostridium

-91<400> 61

<td>agagtttgat</td><td>cctggctcag</td><td>gatgaacgct</td><td>ggcggcgtgc</td><td>ttaacacatg</td><td>caagtcgaac</td><td>60</td>

<td>ggagatatca</td><td>ttttcgaagc</td><td>gattagttta</td><td>ctaagagcgg</td><td>agatgttgct</td><td>atcttagtgg</td><td>120</td>

<td>cggacgggtg</td><td>agtaacgcgt</td><td>gggtaacctg</td><td>ccttgcactg</td><td>ggggataaca</td><td>cttagaaata</td><td>180</td>

<td>ggtgctaata</td><td>ccgcataaca</td><td>gtaggagacg</td><td>catgtctttt</td><td>acttgaaaac</td><td>tccggtggtg</td><td>240</td>

<td>taagatggac</td><td>ccgcgtctga</td><td>ttagcttgtt</td><td>ggcggggtaa</td><td>cggcccacca</td><td>aggcaacgat</td><td>300</td>

<td>cagtagccgg</td><td>cctgagaggg</td><td>tgaacggcca</td><td>cattgggact</td><td>gagacacggc</td><td>ccaaactcct</td><td>360</td>

<td>acgggaggca</td><td>gcagtgggga</td><td>atattggaca</td><td>atggggggaa</td><td>ccctgatcca</td><td>gcgacgccgc</td><td>420</td>

<td>gtgagtgaag</td><td>aagtatttcg</td><td>gtatgtaaag</td><td>ctctatcagc</td><td>agggaagaaa</td><td>gaaatgacgg</td><td>480</td>

<td>tacctgacta</td><td>agaagccccg</td><td>gctaactacg</td><td>tgccagcagc</td><td>cgcggtaata</td><td>cgtagggggc</td><td>540</td>

<td>aagcgttatc</td><td>cggatttact</td><td>gggtgtaaag</td><td>ggagcgtaga</td><td>cggcgatgca</td><td>agtctgaagt</td><td>600</td>

<td>gaaaggcggg</td><td>ggcccaaccc</td><td>ccggactgct</td><td>ttggaaactg</td><td>tatggctgga</td><td>gtgcaggaga</td><td>660</td>

<td>ggtaagtgga</td><td>attcctagtg</td><td>tagcggtgaa</td><td>atgcgtagat</td><td>attaggagga</td><td>acaccagtgg</td><td>720</td>

<td>cgaaagcggc</td><td>ttactggact</td><td>gtaactgacg</td><td>ttgaggctcg</td><td>aaagcgtggg</td><td>gagcaaacaa</td><td>780</td>

<td>gattagatac</td><td>ctggtagtca</td><td>cgccgtaaac</td><td>gatgatcacc</td><td>ggtttcggtg</td><td>ggttatggac</td><td>840</td>

<td>ccatcggttg</td><td>cgcagcaaac</td><td>gcagtagtga</td><td>tccacctggg</td><td>gagtaacgtt</td><td>cgcaagaatg</td><td>900</td>

<td>aaacttcaaa</td><td>ggaaatgacg</td><td>ggggacccgg</td><td>cacaagcggt</td><td>ggaggcatgt</td><td>gtttaattcg</td><td>960</td>

<td>aagcaacgcg</td><td>aagaacctta</td><td>cccaagtctt</td><td>gacatcccgt</td><td>gacgagtgag</td><td>taacgtcact</td><td>1020</td>

<td>ttcccttcgg</td><td>ggcagcggag</td><td>acaggtggtg</td><td>catggttgtc</td><td>gtcagctcgt</td><td>gtcgtgagat</td><td>1080</td>

<td>gttgggttaa</td><td>gtcccgcaac</td><td>gagcgcaacc</td><td>cctatcctta</td><td>gtagccagcg</td><td>agttaggtcg</td><td>1140</td>

<td>ggcactctag</td><td>ggagactgcc</td><td>ggggacaacc</td><td>cggaggaagg</td><td>tggggatgac</td><td>gtcaaatcat</td><td>1200</td>

<td>catgcccctt</td><td>atgatttggg</td><td>ctacacacgt</td><td>gctacaatgg</td><td>cgtaaacaaa</td><td>gggaagcgag</td><td>1-260</td>

<td>cctgtgaagg</td><td>taagcgaatc</td><td>ccagaaataa</td><td>cgtctcagtt</td><td>cggattgtag</td><td>tctgcaactc</td><td>1320</td>

<td>gactacatga</td><td>agctggaatc</td><td>gctagtaatc</td><td>gcggatcaga</td><td>atgccgcggt</td><td>gaatacgttc</td><td>1380</td>

<td>ccgggtcttg</td><td>tacacaccgc</td><td>ccgtcacacc</td><td>atgggagtcg</td><td>gaaatgcccg</td><td>aagtctgtga</td><td>1440</td>

<td>cccaacctga</td><td>gaaggaggga</td><td>gcagccgaag</td><td>gcaggtcgga</td><td>tgactggggt</td><td>gaagtcgtaa</td><td>1500</td>

<td>caaggtagcc</td><td>gt</td><td></td><td></td><td></td><td></td><td>1512</td>

<210> 62 <211> 17 <212> DNA <213> Sztuczna <220>

<223> Sztucznie syntetyzowana sekwencja startera <400> 62 ggtgaatacg ttcccgg 17 <210> 63 <211> 22

-92<212> DNA <213> Sztuczna <220>

<223> Sztucznie syntetyzowana sekwencja startera <400> 63 tacggctacc ttgttacgac tt 22 <210> 64 <211> 20 <212> DNA <213> Sztuczna <220>

<223> Sztucznie syntetyzowana sekwencja startera <400> 64 aaatgacggt acctgactaa 20 <210> 65 <211> 22 <212> DNA <213> Sztuczna <220>

<223> Sztucznie syntetyzowana sekwencja startera <400> 65 ctttgagttt cattcttgcg aa 22 <210> 66 <211> 16 <212> DNA <213> Sztuczna <220>

<223> Sztucznie syntetyzowana sekwencja startera <400> 66 gcacaagcag tggagt 16 <210> 67 <211> 18 <212> DNA <213> Sztuczna <220>

-93<223> Sztucznie syntetyzowana sekwencja startera <400> 67 cttcctccgt tttgtcaa 18 <210> 68 <211> 18 <212> DNA <213> Sztuczna <220>

<223> Sztucznie syntetyzowana sekwencja startera <400> 68 gagaggaagg tcccccac 18 <210> 69 <211> 19 <212> DNA <213> Sztuczna <220>

<223> Sztucznie syntetyzowana sekwencja startera <400> 69 cgctacttgg ctggttcag 19

Piotr Godlewski Rzecznik patentowy