AU723397B2 - Recombinant toxin fragments

48 claims: 10 independent, 38 dependent

1
115 CLAIMS 1. A polypeptide comprising first and second domains, wherein said first domain is adapted to cleave one or more vesicle or plasmamembrane associated proteins essential to exocytosis, said second domain is adapted (i) to translocate the polypeptide into a cell or (ii) to increase the solubility of the polypeptide compared to the solubility of the first domain on its own or (iii) both to translocate the polypeptide into a cell and to increase the solubility of the polypeptide compared to the solubility of the first domain on its own, said polypeptide is free of clostridial neurotoxin and free of clostridial neurotoxin precursor that can be converted into toxin by proteolytic action, and • ·· said polypeptide is a single polypeptide. • · · · • · · “ • · · .·*··.
7
A polypeptide according to any of Claims 1 -5 wherein said second domain clostridial toxin heavy chain H N portion. 116
9
A polypeptide according to any of Claims 1-8 further comprising a third domain adapted for binding of the polypeptide to a cell, by binding of the third domain directly to a cell or by binding of the third domain to a ligand or to ligands that bind to a cell.
16
18. A polypeptide according to any of Claims 1-13 comprising a botulinum toxin light chain or a fragment or a v’ariant of a botulinum toxin light chain and at least 100 N-terminal amino acids of a botulinum toxin heavy chain.
23
25. A polypeptide according to any of Claims 14-24 lacking a portion designated of a botulinum toxin heavy chain. A ft) 118
28
30. A fusion protein comprising a fusion of (a) a polypeptide according to any j*”*· of Claims 1-29 with (b) a second polypeptide being a polypeptide or oligopeptide adapted for binding to an affinity matrix so as to enable purification of the fusion ·«·· :Γ: protein using said matrix. ·«·· j 31 a fusion protein according to Claim 30 wherein said second polypeptide is CM adapted to bind to a chromatography column, such as an affinity matrix of glutathione Sepharose. 119
35
40. A nucleic acid according to Claim 37 and comprising nucleotides encoding residues 1-470 of a botulinum toxin type B light chain.
36
41. A nucleic acid according to Claim 37 or 40 comprising nucleotides encoding residues 1-417 of a botulinum toxin type B heavy chain H N domain.
37
42. A nucleic acid according to any of Claims 37-41 comprising nucleotides encoding a restriction endonuclease cleavage site not present in native clostridial toxin sequence.
39
44. A DNA according to any of claims 37-43.
40
45. A DNA selected from SEQ ID No:s 1,8, 10, 12, 14, 16, 18, 23 and 24.
41
46. A method of manufacture of a polypeptide according to any of Claims 1-29 comprising expressing in a host cell a nucleic acid according to any of Claims^ 3745 and recovering the polypeptide. 4 7 A method of manufacture of a polypeptide according to any of Claims comprising expressing in a host cell a nucleic acid encoding a fusion protein according to Claim 30, 31 or 32 purifying the fusion protein by eluting the fusion protein through an affinity matrix adapted to retain the fusion protein and eluting through said matrix a ligand adapted to displace the fusion protein, and recovering the fusion protein. 121
42
48. A method of manufacture according to Claims 46 or 47 in which the nucleic acid is DNA.
43
49. A cell expressing a polypeptide or fusion protein according to any of 5 Claims 1-32.

Independent claims14

3,380 paragraphs in 714 sections, as filed

WO 98/07864 PCT/GB97/02273

RECOMBINANT TOXIN FRAGMENTS

This invention relates to recombinant toxin fragments, to DNA encoding these fragments and to their uses such as in a vaccine and for in vitro and in vivo purposes.

The clostridial neurotoxins are potent inhibitors of calcium-dependent neurotransmitter secretion in neuronal cells. They are currently considered to mediate this activity through a specific endoproteolytic cleavage of at least one of three vesicle or pre-synaptic membrane associated proteins VAMP, syntaxin or SNAP-25 which are central to the vesicle docking and membrane fusion events of neurotransmitter secretion. The neuronal cell targeting of tetanus and botulinum neurotoxins is considered to be a receptor mediated event following which the toxins become internalised and subsequently traffic to the appropriate intracellular compartment where they effect their endopeptidase activity.

The clostridial neurotoxins share a common architecture of a catalytic L-chain (LC, ca 50 kDa) disulphide linked to a receptor binding and translocating H-chain (HC, ca 100 kDa). The HC polypeptide is considered to comprise all or part of two distinct functional domains. The carboxy-terminal half of the HC (ca 50 kDa), termed the Hc domain, is involved in the high affinity, neurospecific binding of the neurotoxin to cell surface receptors on the target neuron, whilst the amino-terminal half, termed the HN domain (ca 50 kDa), is considered to mediate the translocation of at least some portion of the neurotoxin across cellular membranes such that the functional activity of the LC is expressed within the target cell. The HN domain also has the property, under conditions of low pH, of forming ion-permeable channels in lipid membranes, this may in some manner relate to its translocation function.

For botulinum neurotoxin type A (BoNT/A) these domains are considered to reside within amino acid residues 872-1296 for the Hc, amino acid residues 449-871 for the Hn and residues 1-448 for the LC. Digestion with trypsin effectively degrades the Hc domain of the BoNT/A to generate a non-toxic fragment designated LHN,

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- 2 which is no longer able to bind to and enter neurons (Fig. 1). The LHN fragment so produced also has the property of enhanced solubility compared to both the parent holotoxin and the isolated LC.

It is therefore possible to provide functional definitions of the domains within the neurotoxin molecule, as follows:

(A) clostridial neurotoxin light chain:

-a metalloprotease exhibiting high substrate specificity for vesicle and/or plasma membrane associated proteins involved in the exocytotic process. In particular, it cleaves one or more of SNAP-25, VAMP (synaptobrevin / cellubrevin) and syntaxin.

(B) clostridial neurotoxin heavy chain HN domain:

-a portion of the heavy chain which enables translocation of that portion of the neurotoxin molecule such that a functional expression of light chain activity occurs within a target cell.

-the domain responsible for translocation of the endopeptidase activity, following binding of neurotoxin to its specific cell surface receptor via the binding domain, into the target cell.

-the domain responsible for formation of ion-permeable pores in lipid membranes under conditions of low pH.

-the domain responsible for increasing the solubility of the entire polypeptide compared to the solubility of light chain alone.

(C) clostridial neurotoxin heavy chain Hc domain.

-a portion of the heavy chain which is responsible for binding of the native

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- 3 holotoxin to cell surface receptor(s) involved in the intoxicating action of clostridial toxin prior to internalisation of the toxin into the cell.

The identity of the cellular recognition markers for these toxins is currently not understood and no specific receptor species have yet been identified although Kozaki et al. have reported that synaptotagmin may be the receptor for botulinum neurotoxin type B. It is probable that each of the neurotoxins has a different receptor.

It is desirable to have positive controls fortoxin assays, to develop clostridial toxin vaccines and to develop therapeutic agents incorporating desirable properties of clostridial toxin.

However, due to its extreme toxicity, the handling of native toxin is hazardous.

The present invention seeks to overcome or at least ameliorate problems associated with production and handling of clostridial toxin.

Accordingly, the invention provides a polypeptide comprising first and second domains, wherein said first domain is adapted to cleave one or more vesicle or plasma-membrane associated proteins essential to neuronal exocytosis and wherein said second domain is adapted (i) to translocate the polypeptide into the cell or (ii) to increase the solubility of the polypeptide compared to the solubility of the first domain on its own or (ill) both to translocate the polypeptide into the cell and to increase the solubility of the polypeptide compared to the solubility of the first domain on its own, said polypeptide being free of clostridial neurotoxin and free of any clostridial neurotoxin precursor that can be converted into toxin by proteolytic action. Accordingly, the invention may thus provide a single polypeptide chain containing a domain equivalent to a clostridial toxin light chain and a domain providing the functional aspects of the HN of a clostridial toxin heavy chain, whilst lacking the functional aspects of a clostridial toxin Hc domain.

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- 4 For the purposes of the invention, the functional property or properties of the HN of a clostridial toxin heavy chain that are required to be exhibited by the second domain of the polypeptide of the invention are either (i) translocation of the polypeptide into a cell, or (ii) increasing solubility of the polypeptide compared to solubility of the first domain on its own or (iii) both (i) and (ii). References hereafter to a Hn domain or to the functions of a HN domain are references to this property or properties. The second domain is not required to exhibit other properties of the Hn domain of a clostridial toxin heavy chain.

A polypeptide of the invention can thus be soluble but lack the translocation function of a native toxin-this is of use in providing an immunogen for vaccinating or assisting to vaccinate an individual against challenge by toxin. In a specific embodiment of the invention described in an example below a polypeptide designated LH423/A elicited neutralising antibodies against type A neurotoxin. A polypeptide of the invention can likewise thus be relatively insoluble but retain the translocation function of a native toxin - this is of use if solubility is imparted to a composition made up of that polypeptide and one or more other components by one or more of said other components.

The first domain of the polypeptide of the invention cleaves one or more vesicle or plasma-membrane associated proteins essential to the specific cellular process of exocytosis, and cleavage of these proteins results in inhibition of exocytosis, typically in a non-cytotoxic manner. The cell or cells affected are not restricted to a particular type or subgroup but can include both neuronal and non-neuronal cells. The activity of clostridial neurotoxins in inhibiting exocytosis has, indeed, been observed almost universally in eukaryotic cells expressing a relevant cell surface receptor, including such diverse cells as from Aplysia (sea slug), Drosophila (fruit fly) and mammalian nerve cells, and the activity of the first domain is to be understood as including a corresponding range of cells.

The polypeptide of the invention may be obtained by expression of a recombinant nucleic acid, preferably a DNA, and is a single polypeptide, that is to say not

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- 5 cleaved into separate light and heavy chain domains. The polypeptide is thus available in convenient and large quantities using recombinant techniques.

In a polypeptide according to the invention, said first domain preferably comprises a clostridial toxin light chain or a fragment or variant of a clostridial toxin light chain. The fragment is optionally an N-terminal, or C-terminal fragment of the light chain, or is an internal fragment, so long as it substantially retains the ability to cleave the vesicle or plasma-membrane associated protein essential to exocytosis. The minimal domains necessary for the activity of the light chain of clostridial toxins are described in J. Biol. Chem., Vol.267, No. 21, July 1 992, pages 1472114729. The variant has a different peptide sequence from the light chain or from the fragment, though it too is capable of cleaving the vesicle or plasma-membrane associated protein. It is conveniently obtained by insertion, deletion and/or substitution of a light chain or fragment thereof. In embodiments of the invention described below a variant sequence comprises (i) an N-terminal extension to a clostridial toxin light chain or fragment (ii) a clostridial toxin light chain or fragment modified by alteration of at least one amino acid (iii) a C-terminal extension to a clostridial toxin light chain or fragment, or (iv) combinations of 2 or more of (i)-(iii).

In further embodiments of the invention, the variant contains an amino acid sequence modified so that (a) there is no protease sensitive region between the LC and Hn components of the polypeptide, or (b) the protease sensitive region is specific for a particular protease. This latter embodiment is of use if it is desired to activate the endopeptidase activity of the light chain in a particular environment or_cell. Though, in general, the polypeptides of the invention are activated prior to administration.

The first domain preferably exhibits endopeptidase activity specific for a substrate selected from one or more of SNAP-25, synaptobrevin/VAMP and syntaxin. The clostridial toxin is preferably botulinum toxin or tetanus toxin.

In an embodiment of the invention described in an example below, the toxin light

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- 6 chain and the portion of the toxin heavy chain are of botulinum toxin type A. In a further embodiment of the invention described in an example below, the toxin light chain and the portion of the toxin heavy chain are of botulinum toxin type B. The polypeptide optionally comprises a light chain or fragment or variant of one toxin type and a heavy chain or fragment or variant of another toxin type.

In a polypeptide according to the invention said second domain preferably comprises a clostridial toxin heavy chain HN portion or a fragment or variant of a clostridial toxin heavy chain HN portion. The fragment is optionally an N-terminal or C-terminal or internal fragment, so long as it retains the function of the HN domain. Teachings of regions within the HN responsible for its function are provided for example in Biochemistry 1995, 34, pages 15175-15181 and Eur. J. Biochem, 1989, 185, pages 197-203. The variant has a different sequence from the Hn domain or fragment, though it too retains the function of the HN domain. It is conveniently obtained by insertion, deletion and/or substitution of a HN domain or fragment thereof. In embodiments of the invention, described below, it comprises (i) an N-terminal extension to a HN domain or fragment, (ii) a C-terminal extension to a HN domain or fragment, (iii) a modification to a HN domain or fragment by alteration of at least one amino acid, or (iv) combinations of 2 or more of (i)-(iii). The clostridial toxin is preferably botulinum toxin or tetanus toxin.

The invention also provides a polypeptide comprising a clostridial neurotoxin light chain and a N-terminal fragment of a clostridial neurotoxin heavy chain, the fragment preferably comprising at least 423 of the N-terminal amino acids of the heavy chain of botulinum toxin type A, 417 of the N-terminal amino acids of the heavy chain of botulinum toxin type B or the equivalent number of N-terminal amino acids of the heavy chain of other types of clostridial toxin such that the fragment possesses an equivalent alignment of homologous amino acid residues.

These polypeptides of the invention are thus not composed of two or more polypeptides, linked for example by di-sulphide bridges into composite molecules. Instead, these polypeptides are single chains and are not active or their activity is

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- 7 significantly reduced in an in vitro assay of neurotoxin endopeptidase activity.

Further, the polypeptides may be susceptible to be converted into a form exhibiting endopeptidase activity by the action of a proteolytic agent, such as trypsin. In this way it is possible to control the endopeptidase activity of the toxin light chain.

In a specific embodiment of the invention described in an example below, there is provided a polypeptide lacking a portion designated Hc of a clostridial toxin heavy chain. This portion, seen in the naturally produced toxin, is responsible for binding of toxin to cell surface receptors prior to internalisation of the toxin. This specific embodiment is therefore adapted so that it can not be converted into active toxin, for example by the action of a proteolytic enzyme. The invention thus also provides a polypeptide comprising a clostridial toxin light chain and a fragment of a clostridial toxin heavy chain, said fragment being not capable of binding to those cell surface receptors involved in the intoxicating action of clostridial toxin, and it is preferred that such a polypeptide lacks an intact portion designated Hc of a clostridial toxin heavy chain.

In further embodiments of the invention there are provided compositions containing a polypeptide comprising a clostridial toxin light chain and a portion designated HN of a clostridial toxin heavy chain, and wherein the composition is free of clostridial toxin and free of any clostridial toxin precursor that may be converted into clostridial toxin by the action of a proteolytic enzyme. Examples of these compositions include those containing toxin light chain and HN sequences of botulinum toxin types A, B, Cv D, E, F and G.

The polypeptides of the invention are conveniently adapted to bind to, or include, a ligand for targeting to desired cells. The polypeptide optionally comprises a sequence that binds to, for example, an immunoglobulin. A suitable sequence is a tandem repeat synthetic IgG binding domain derived from domain B of Staphylococcal protein A. Choice of immunoglobulin specificity then determines the target for a polypeptide - immunoglobulin complex. Alternatively, the

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- 8 polypeptide comprises a non-clostridial sequence that binds to a cell surface receptor, suitable sequences including insulin-like growth factor-1 (IGF-1) which binds to its specific receptor on particular cell types and the 14 amino acid residue sequence from the carboxy-terminus of cholera toxin A subunit which is able to bind the cholera toxin B subunit and thence to GM1 gangliosides. A polypeptide according to the invention thus, optionally, further comprises a third domain adapted for binding of the polypeptide to a cell.

In a second aspect the invention provides a fusion protein comprising a fusion of (a) a polypeptide of the invention as described above with (b) a second polypeptide adapted for binding to a chromatography matrix so as to enable purification of the fusion protein using said chromatography matrix. It is convenient for the second polypeptide to be adapted to bind to an affinity matrix, such as a glutathione Sepharose, enabling rapid separation and purification of the fusion protein from an impure source, such as a cell extract or supernatant.

One possible second purification polypeptide is glutathione-S-transferase (GST), and others will be apparent to a person of skill in the art, being chosen so as to enable purification on a chromatography column according to conventional techniques.

As noted above, by proteolytic treatment, for example using trypsin, of a polypeptide of the invention it is possible to induce endopeptidase activity in the treated polypeptide. A third aspect of the invention provides a composition comprising a derivative of a clostridial toxin, said derivative retaining at least 10% of the endopeptidase activity of the clostridial toxin, said derivative further being non-toxic in vivo due to its inability to bind to cell surface receptors, and wherein the composition is free of any component, such as toxin or a further toxin derivative, that is toxic in vivo. The activity of the derivative preferably approaches that of natural toxin, and is thus preferably at least 30% and most preferably at least 60% of natural toxin. The overall endopeptidase activity of the composition will, of course, also be determined by the amount of the derivative that is present.

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- 9 While it is known to treat naturally produced clostridial toxin to remove the Hc domain, this treatment does not totally remove toxicity of the preparation, instead some residual toxin activity remains. Natural toxin treated in this way is therefore still not entirely safe. The composition of the invention, derived by treatment of a pure source of polypeptide advantageously is free of toxicity, and can conveniently be used as a positive control in a toxin assay, as a vaccine against clostridial toxin or for other purposes where it is essential that there is no residual toxicity in the composition.

The invention enables production of the polypeptides and fusion proteins of the invention by recombinant means.

A fourth aspect of the invention provides a nucleic acid encoding a polypeptide or a fusion protein according to any of the aspects of the invention described above.

In one embodiment of this aspect of the invention, a DNA sequence provided to code for the polypeptide or fusion protein is not derived from native clostridial sequences, but is an artificially derived sequence not preexisting in nature.

A specific DNA (SEQ ID NO: 1) described in more detail below encodes a polypeptide or a fusion protein comprising nucleotides encoding residues 1-871 of a botulinum toxin type A. Said polypeptide comprises the light chain domain and the first 423 amino acid residues of the amino terminal portion of a botulinum toxin type A heavy chain. This recombinant product is designated LH423/A (SEQ ID NO: 2). .

In a second embodiment of this aspect of the invention a DNA sequence which codes for the polypeptide or fusion protein is derived from native clostridial sequences but codes for a polypeptide or fusion protein not found in nature.

A specific DNA (SEQ ID NO: 19) described in more detail below encodes a polypeptide or a fusion protein and comprises nucleotides encoding residues 1WO 98/07864

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- 10 1171 of a botulinum toxin type B. Said polypeptide comprises the light chain domain and the first 728 amino acid residues of the amino terminal protein of a botulinum type B heavy chain. This recombinant product is designated LH728/B (SEQ ID NO: 20).

The invention thus also provides a method of manufacture of a polypeptide comprising expressing in a host cell a DNA according to the third aspect of the invention. The host cell is suitably not able to cleave a polypeptide or fusion protein of the invention so as to separate light and heavy toxin chains; for example, a non-clostridial host.

The invention further provides a method of manufacture of a polypeptide comprising expressing in a host cell a DNA encoding a fusion protein as described above, purifying the fusion protein by elution through a chromatography column adapted to retain the fusion protein, eluting through said chromatography column a ligand adapted to displace the fusion protein and recovering the fusion protein. Production of substantially pure fusion protein is thus made possible. Likewise, the fusion protein is readily cleaved to yield a polypeptide of the invention, again in substantially pure form, as the second polypeptide may conveniently be removed using the same type of chromatography column.

The LHn/A derived from dichain native toxin requires extended digestion with trypsin to remove the C-terminal 1/2 of the heavy chain, the Hc domain. The loss of this domain effectively renders the toxin inactive in vivo by preventing its interaction with host target cells. There is, however, a residual toxic activity which may indicate a contaminating, trypsin insensitive, form of the whole type A neurotoxin.

In contrast, the recombinant preparations of the invention are the product of a discreet, defined gene coding sequence and can not be contaminated by full length toxin protein. Furthermore, the product as recovered from E. coli, and from other recombinant expression hosts, is an inactive single chain peptide or if expression

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- 11 hosts produce a processed, active polypeptide it is not a toxin. Endopeptidase activity of LH423/A, as assessed by the current in vitro peptide cleavage assay, is wholly dependent on activation of the recombinant molecule between residues 430 and 454 by trypsin. Other proteolytic enzymes that cleave between these two residues are generally also suitable for activation of the recombinant molecule. Trypsin cleaves the peptide bond C-terminal to Arginine or C-terminal to Lysine and is suitable as these residues are found in the 430-454 region and are exposed (see Fig. 12).

The recombinant polypeptides of the invention are potential therapeutic agents for targeting to cells expressing the relevant substrate but which are not implicated in effecting botulism. An example might be where secretion of neurotransmitter is inappropriate or undesirable or alternatively where a neuronal cell is hyperactive in terms of regulated secretion of substances other than neurotransmitter. In such an example the function of the Hc domain of the native toxin could be replaced by an alternative targeting sequence providing, for example, a cell receptor ligand and/or translocation domain.

One application of the recombinant polypeptides of the invention will be as a reagent component for synthesis of therapeutic molecules, such as disclosed in WO-A-94/21300. The recombinant product will also find application as a non-toxic standard for the assessment and development of in vitro assays for detection of functional botulinum or tetanus neurotoxins either in foodstuffs or in environmental samples, for example as disclosed in EP-A-0763131.

A further option is addition, to the C-terminal end of a polypeptide of the invention, of a peptide sequence which allows specific chemical conjugation to targeting ligands of both protein and non-protein origin.

In yet a further embodiment an alternative targeting ligand is added to the Nterminus of polypeptides of the invention. Recombinant LHN derivatives have been designated that have specific protease cleavage sites engineered at the C-terminus

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- 12 of the LC at the putative trypsin sensitive region and also at the extreme Cterminus of the complete protein product. These sites will enhance the activational specificity of the recombinant product such that the dichain species can only be activated by proteolytic cleavage of a more predictable nature than use of trypsin.

The LHn enzymatically produced from native BoNT/A is an efficient immunogen and thus the recombinant form with its total divorce from any full length neurotoxin represents a vaccine component. The recombinant product may serve as a basal reagent for creating defined protein modifications in support of any of the above areas.

Recombinant constructs are assigned distinguishing names on the basis of their amino acid sequence length and their Light Chain (L-chain, L) and Heavy Chain (Hchain, H) content as these relate to translated DNA sequences in the public domain or specifically to SEQ ID NO: 2 and SEQ ID NO: 20. The 'LH' designation is followed by 7X' where 'X' denotes the corresponding clostridial toxin serotype or class, e.g. A for botulinum neurotoxin type A or 'TeTx' for tetanus toxin. Sequence variants from that of the native toxin polypeptide are given in parenthesis in standard format, namely the residue position number prefixed by the residue of the native sequence and suffixed by the residue of the variant.

Subscript number prefixes indicate an amino-terminal (N-terminal) extension, or where negative a deletion, to the translated sequence. Similarly, subscript number suffixes indicate a carboxy terminal (C-terminal) extension or where negative numbers are used, a deletion. Specific sequence inserts such as protease cleavage sites are indicated using abbreviations, e.g. Factor Xa is abbreviated to FXa. Lchain C-terminal suffixes and H-chain N-terminal prefixes are separated by a 7' to indicate the predicted junction between the L and H-chains. Abbreviations for engineered ligand sequences are prefixed or suffixed to the clostridial L-chain or Hchain corresponding to their position in the translation product.

Following this nomenclature,

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<td> LH423/A</td><td> = SEQ ID NO: 2, containing the entire L-chain and 423 amino acids of the H-chain of botulinum neurotoxin type A;</td>

<td> 2LH423/A</td><td> a variant of this molecule, containing a two amino acid</td>

<td></td><td> extension to the N-terminus of the L-chain;</td>

<td> 2L/2H423/A</td><td> = a further variant in which the molecule contains a two amino acid extension on the N-terminus of both the Lchain and the H-chain;</td>

<td> 2^~FXa/2^423^A</td><td> = a further variant containing a two amino acid extension to the N-terminus of the L-chain, and a Factor Xa cleavage sequence at the C-terminus of the L-chain which, after cleavage of the molecule with Factor Xa leaves a two amino acid N-terminal extension to the Hchain component; and</td>

<td> 2L-FXa/2^423/A-IGF-1</td><td> = a variant of this molecule which has a further C-terminal extension to the H-chain, in this example the insulin-like growth factor 1 (IGF-1) sequence.</td>

There now follows description of specific embodiments of the invention, illustrated by drawings in which:

<td> Fig. 1</td><td> shows a schematic representation of the domain structure of botulinum neurotoxin type A (BoNT/A);</td>

<td> Fig. 2</td><td> shows a schematic representation of assembly of the gene for an embodiment of the invention designated LH423/A;</td>

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<td> Fig. 3</td><td> is a graph comparing activity of native toxin, trypsin generated native LHN/A and an embodiment of the invention designated 2LH423/A (α2Ε,Ν26Κ,Α27Υ) in an in vitro peptide cleavage assay;</td>

<td> Fig. 4</td><td> is a comparison of the first 33 amino acids in published sequences of native toxin and embodiments of the invention;</td>

<td> Fig. 5</td><td> shows the transition region of an embodiment of the invention designated L/4H423/A illustrating insertion of four amino acids at the N-terminus of the HN sequence; amino acids coded for by the Eco 47 III restriction endonuclease cleavage site are marked and the HN sequence then begins ALN...;</td>

<td> Fig. 6</td><td> shows the transition region of an embodiment of the invention designated LFXa/3H423/A illustrating insertion of a Factor Xa cleavage site at the C-terminus of the L-chain, and three additional amino acids coded for at the N-terminus of the Flsequence; the N-terminal amino acid of the cleavage-activated Hn will be cysteine;</td>

<td> Fig. 7</td><td> shows the C-terminal portion of the amino acid sequence of an embodiment of the invention designated LFXa/3H423/A-IGF-1, a fusion protein; the IGF-1 sequence begins at position G882;</td>

<td> Fig. 8</td><td> shows the C-terminal portion of the amino acid sequence of an embodiment of the invention designated LFXa/3H423/A-CtxA14, a fusion protein; the C-terminal CtxA sequence begins at position Q882;</td>

<td> Fig.9</td><td> shows the C-terminal portion of the amino acid sequence ofan</td>

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<td></td><td> - 15 - embodiment of the invention designated LFXa/3H423/A-ZZ, a fusion protein; the C-terminal ZZ sequence begins at position A890 immediately after a genenase recognition site (underlined);</td>

show schematic representations of manipulations of

Figs. 10 & 11 polypeptides of the invention; Fig. 10 shows LH423/A

<td></td><td> with N-terminal addition of an affinity purification peptide (in this case GST) and C-terminal addition of an lg binding domain; protease cleavage sites R1, R2 and R3 enable selective enzymatic separation of domains; Fig. 11 shows specific examples of protease cleavage sites R1, R2 and R3 and a C-terminal fusion peptide sequence;</td>

<td> Fig. 12</td><td> shows the trypsin sensitive activation region of a polypeptide of the invention;</td>

<td> Fig. 13</td><td> shows Western blot analysis of recombinant LH107/B expressed from E.colr, panel A was probed with anti-</td>

<td></td><td> BoNT/B antiserum; Lane 1, molecular weight standards; lanes 2 & 3, native BoNT/B; lane 4, immunopurified LH107/B; panel B was probed with anti-T7 peptide tag antiserum; lane 1, molecular weight standards; lanes 2 & 3, positive control E.coH T7 expression; lane 4 immunopurified LH107/B.</td>

The sequence listing that accompanies this application contains the following sequences:-

<td> SEQ ID NO:</td><td> Sequence</td>

<td> 1</td><td> DNA coding for LH423/A</td>

<td> 3</td><td> DNA coding for 23l-H423/A (Q2E,N26K,A27Y), of which an N-terminal portion is shown in Fig. 4.</td>

<td> 5</td><td> DNA coding for 2LH423/A (Q2E,N26K,A27Y), of which an Nterminal portion is shown in Fig.4</td>

<td> 7</td><td> DNA coding for native BoNT/A according to Binz et al</td>

<td> 8</td><td> native BoNT/A according to Binz et al</td>

<td> 9</td><td> DNA coding for L/4H4237A</td>

<td> 11</td><td> DNA coding for LFXa/3H423/A</td>

<td> 13</td><td> DNA coding for LFXa/3H423/A-IGF-1</td>

<td> 15</td><td> DNA coding for LFXa/3H423/A-CtxA14</td>

<td> 17</td><td> DNA coding for LFXa/3H423/A-ZZ</td>

<td> 19</td><td> DNA coding for LH728/B</td>

<td> 21</td><td> DNA coding for LH417/B</td>

<td> 23</td><td> DNA coding for LH107/B</td>

<td> 25;</td><td> DNA coding for LH423/A (Q2E,N26K,A27Y)</td>

<td> 27</td><td> DNA coding for LH417/B wherein the first 274 bases are</td>

- 17 28 modified to have an E.co/i codon bias

DNA coding for LH417/B wherein bases 691-1641 of the native BoNT/B sequence have been replaced by a degenerate DNA coding for amino acid residues 231 -547 of the native BoNT/B polypeptide

In the claims which follow and in the preceding summary of the invention, except where the context requires otherwise due to express language or necessary implication, the word comprising is used in the sense of including, i.e. the features specified may be associated with further features in various embodiments of the invention.

Example 1

A 2616 base pair, double stranded gene sequence (SEQ ID NO: 1) has been assembled from a combination of synthetic, chromosomal and polymerase-chain-reaction generated DNA (Figure 2). The gene codes for a polypeptide of 871 ammo acid residues corresponding to the entire light-chain (LC, 448 ammo acids) and 423 residues of the amino terminus of the heavy-chain (Hc) of botulinum neurotoxin type A. This recombinant product is designated the LH423/A fragment (SEQ ID NO: 2). 43 j Construction of the recombinant product • · · • •4 · ····

4 ·

...... The first 918 base pairs of the recombinant gene were synthesised by concatenation of short oligonucleotides to generate a coding sequence with an E. col/ codon bias. Both DNA strands in this region were completely synthesised as short overlapping oligonucleotides which were phosphorylated, annealed and H9ated tO generate the ful1 synthetic region ending with a unique Kpn\ restriction S,te' The remainder of LH423/A coding sequence was PCR amplified from total chromosomal DNA from Clostridium botulinum and annealed to the synthetic ί ♦ portion of the gene. 4 »

:. The internal PCR amplified product sequences were then deleted and replaced with _ the native, fully sequenced, regions from clones of C. botulinum chromosomal origin to generate the final gene construct. The final composition is synthetic DNA ^AR^toases 1-913), polymerase amplified DNA (bases 914-1138 and 1976-2616) and

-o u tf rema’nder ,s of C’ botulinum chromosomal origin (bases 1139-1975) The <£yj ’

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- 18 assembled gene was then fully sequenced and cloned into a variety of E.coli plasmid vectors for expression analysis.

Expression of the recombinant gene and recovery of protein product

The DNA is expressed in E. coli as a single nucleic acid transcript producing a soluble single chain polypeptide of 99,951 Daltons predicted molecular weight. The gene is currently expressed in E. coli as a fusion to the commercially available coding sequence of glutathione S-transferase (GST) of Schistosoma japonicum but any of an extensive range of recombinant gene expression vectors such as pEZZ18, pTrc99, pFLAG or the pMAL series may be equally effective as might expression in other prokaryotic or eukaryotic hosts such as the Gram positive bacilli, the yeast P. pastoris or in insect or mammalian cells under appropriate conditions.

Currently, E. coli harbouring the expression construct is grown in Luria-Bertani broth (L-broth pH 7.0, containing 10 g/l bacto-tryptone, 5 g/l bacto-yeast extract and 10 g/l sodium chloride) at 37° C until the cell density (biomass) has an optical absorbance of 0.4- 0.6 at 600 nm and the cells are in mid-logarithmic growth phase. Expression of the gene is then induced by addition of isopropylthio-jff-D-galactosidase (IPTG) to a final concentration of 0.5 mM. Recombinant gene expression is allowed to proceed for 90 min at a reduced temperature of 25°C. The cells are then harvested by centrifugation, are resuspended in a buffer solution containing 10 mM Na2HP04, 0.5 M NaCl, 10 mM EGTA, 0.25% Tween, pH 7.0 and then frozen at -20°C. For extraction of the recombinant protein the cells are_disrupted by sonication. The cell extract is then cleared of debris by centrifugation and the cleared supernatant fluid containing soluble recombinant fusion protein (GST- LH423/A) is stored at -20°C pending purification. A proportion of recombinant material is not released by the sonication procedure and this probably reflects insolubility or inclusion body formation. Currently we do not extract this material for analysis but if desired this could be readily achieved using methods known to those skilled in the art.

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- 19 The recombinant GST- LH423/A is purified by adsorption onto a commercially prepared affinity matrix of glutathione Sepharose and subsequent elution with reduced glutathione. The GST affinity purification marker is then removed by proteolytic cleavage and reabsorption to glutathione Sepharose; recombinant LH423/A is recovered in the non-adsorbed material.

Construct variants

A variant of the molecule, LH423/A (Q2E,N26K,A27Y) (SEQ ID NO: 26) has been produced in which three amino acid residues have been modified within the light chain of LH423/A producing a polypeptide containing a light chain sequence different to that of the published amino acid sequence of the light chain of BoNT/A .

Two further variants of the gene sequence that have been expressed and the corresponding products purified are 23LH423/A (Q2E,N26K,A27Y) (SEQ ID NO: 4) which has a 23 amino acid N-terminal extension as compared to the predicted native L-chain of BoNT/A and 2LH423/A (Q2E,N26K,A27Y) (SEQ ID NO: 6) which has a 2 amino acid N-terminal extension (Figure 4).

In yet another variant a gene has been produced which contains a Eco 47 III restriction site between nucleotides 1344 and 1345 of the gene sequence given in (SEQ ID NO: 1). This modification provides a restriction site at the position in the gene representing the interface of the heavy and light chains in native neurotoxin, and provides the capability to make insertions at this point using standard restriction enzyme methodologies known to those skilled in the art. It will also be obvious to those skilled in the art that any one of a number of restriction sites could be so employed, and that the Eco 47 III insertion simply exemplifies this approach. Similarly, it would be obvious for one skilled in the art that insertion of a restriction site in the manner described could be performed on any gene of the invention. The gene described, when expressed, codes for a polypeptide, L/4H423/A (SEQ ID NO: 10), which contains an additional four amino acids between amino acids 448 and 449 of LH423/A at a position equivalent to the amino terminus of the

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- 20 heavy chain of native BoNT/A.

A variant of the gene has been expressed, LFXa/3H423/A (SEQ ID NO: 12), in which a specific proteolytic cleavage site was incorporated at the carboxy-terminal end of the light chain domain, specifically after residue 448 of L/4H423/A. The cleavage site incorporated was for Factor Xa protease and was coded for by modification of SEQ ID NO: 1. It will be apparent to one skilled in the art that a cleavage site for another specified protease could be similarly incorporated, and that any gene sequence coding for the required cleavage site could be employed. Modification of the gene sequence in this manner to code for a defined protease site could be performed on any gene of the invention.

Variants of LFXa/3H423/A have been constructed in which a third domain is present at the carboxy-terminal end of the polypeptide which incorporates a specific binding activity into the polypeptide.

Specific examples described are:

(1) LFxa/3H423/A-IGF-1 (SEQ ID NO: 14) , in which the carboxy-terminal domain has a sequence equivalent to that of insulin-like growth factor-1 (IGF-1) and is able to bind to the insulin-like growth factor receptor with high affinity;

(2) LFXa/3H423/A-CtxA14 (SEQ ID NO: 16) , in which the carboxy-terminal domain has a sequence equivalent to that of the 14 amino acids from the carboxy-terminus of the A-subunit of cholera toxin (CtxA) and is thereby able to interact with the cholera toxin B-subunit pentamer,· and (3) LFXa/3H423/A-ZZ (SEQ ID NO: 18) , in which the carboxy-terminal domain is a tandem repeating synthetic IgG binding domain. This variant also exemplifies another modification applicable to the current invention, namely the inclusion in the gene of a sequence coding for a protease cleavage site located between the end of the clostridial heavy chain sequence and the sequence coding for the binding

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- 21 ligand. Specifically in this example a sequence is inserted at nucleotides 2650 to 2666 coding for a genenase cleavage site. Expression of this gene produces a polypeptide which has the desired protease sensitivity at the interface between the domain providing HN function and the binding domain. Such a modification enables selective removal of the C-terminal binding domain by treatment of the polypeptide with the relevant protease.

It will be apparent that any one of a number of such binding domains could be incorporated into the polypeptide sequences of this invention and that the above examples are merely to exemplify the concept. Similarly, such binding domains can be incorporated into any of the polypeptide sequences that are the basis of this invention. Further, it should be noted that such binding domains could be incorporated at any appropriate location within the polypeptide molecules of the invention.

Further embodiments of the invention are thus illustrated by a DNA of the invention further comprising a desired restriction endonuclease site at a desired location and by a polypeptide of the invention further comprising a desired protease cleavage site at a desired location.

The restriction endonuclease site may be introduced so as to facilitate further manipulation of the DNA in manufacture of an expression vector for expressing a polypeptide of the invention; it may be introduced as a consequence of a previous step in manufacture of the DNA; it may be introduced by way of modification by insertion, substitution or deletion of a known sequence. The consequence of modification of the DNA may be that the amino acid sequence is unchanged, or may be that the amino acid sequence is changed, for example resulting in introduction of a desired protease cleavage site, either way the polypeptide retains its first and second domains having the properties required by the invention.

Figure 10 is a diagrammatic representation of an expression product exemplifying features described in this example. Specifically, it illustrates a single polypeptide

WO 98/07864 PCT/GB97/02273

- 22 incorporating a domain equivalent to the light chain of botulinum neurotoxin type A and a domain equivalent to the HN domain of the heavy chain of botulinum neurotoxin type A with a N-terminal extension providing an affinity purification domain, namely GST, and a C-terminal extension providing a ligand binding domain, namely an IgG binding domain. The domains of the polypeptide are spatially separated by specific protease cleavage sites enabling selective enzymatic separation of domains as exemplified in the Figure. This concept is more specifically depicted in Figure 11 where the various protease sensitivities are defined for the purpose of example.

Assay of product activity

The LC of botulinum neurotoxin type A exerts a zinc-dependent endopeptidase activity on the synaptic vesicle associated protein SNAP-25 which it cleaves in a specific manner at a single peptide bond. The 2LH423/A (Q2E,N26K,A27Y) (SEQ ID NO: 6) cleaves a synthetic SNAP-25 substrate in vitro under the same conditions as the native toxin (Figure 3). Thus, the modification of the polypeptide sequence of 2LH423/A (Q2E,N26K,A27Y) relative to the native sequence and within the minimal functional LC domains does not prevent the functional activity of the LC domains.

This activity is dependent on proteolytic modification of the recombinant GST-2LH423/A (Q2E,N26K,A27Y) to convert the single chain polypeptide product to a disulphide linked dichain species. This is currently done using the proteolytic enzyme trypsin. The recombinant product (100-600 pg/ml) is incubated at 37°C for 10-50 minutes with trypsin (10pg/ml) in a SQ.Iution containing 140 mM NaCI,

2.7 mM KCI, 10 mM Na2HP04, 1.8 mM KH2PO4, pH 1.3. The reaction is terminated by addition of a 100-fold molar excess of trypsin inhibitor. The activation by trypsin generates a disulphide linked dichain species as determined by polyacrylamide gel electrophoresis and immunoblotting analysis using polyclonal anti-botulinum neurotoxin type A antiserum.

2LH423/A is more stable in the presence of trypsin and more active in the in vitro

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- 23 peptide cleavage assay than is 23LH423/A. Both variants, however, are fully functional in the in vitro peptide cleavage assay. This demonstrates that the recombinant molecule will tolerate N-terminal amino acid extensions and this may be expanded to other chemical or organic moieties as would be obvious to those skilled in the art.

Example 2

As a further exemplification of this invention a number of gene sequences have been assembled coding for polypeptides corresponding to the entire light-chain and varying numbers of residues from the amino terminal end of the heavy chain of botulinum neurotoxin type B. In this exemplification of the disclosure the gene sequences assembled were obtained from a combination of chromosomal and polymerase-chain-reaction generated DNA, and therefore have the nucleotide sequence of the equivalent regions of the natural genes, thus exemplifying the principle that the substance of this disclosure can be based upon natural as well as a synthetic gene sequences.

The gene sequences relating to this example were all assembled and expressed using methodologies as detailed in Sambrook J, Fritsch E F & Maniatis T (1989) Molecular Cloning: A Laboratory Manual (2nd Edition), Ford N, Nolan C, Ferguson M & Ockler M (eds), Cold Spring Harbor Laboratory Press, New York, and known to those skilled in the art.

A_ gene has been assembled coding for a polypeptide of 1171 amino acids corresponding to the entire light-chain (443 amino acids) and 728 residues from the amino terminus of the heavy chain of neurotoxin type B. Expression of this gene produces a polypeptide, LH72g/B (SEQ ID NO: 20), which lacks the specific neuronal binding activity of full length BoNT/B.

A gene has also been assembled coding for a variant polypeptide, LH417/B (SEQ ID NO: 22), which possesses an amino acid sequence at its carboxy terminus

WO 98/07864 PCT/GB97/02273

- 24 equivalent by amino acid homology to that at the carboxy-terminus of the heavy chain fragment in native LHN/A .

A gene has also been assembled coding for a variant polypeptide, LH107/B (SEQ ID NO: 24) , which expresses at its carboxy-terminus a short sequence from the amino terminus of the heavy chain of BoNT/B sufficient to maintain solubility of the expressed polypeptide.

Construct Variants

A variant of the coding sequence for the first 274 bases of the gene shown in SEQ ID NO: 21 has been produced which whilst being a non-native nucleotide sequence still codes for the native polypeptide.

Two double stranded, a 268 base pair and a 951 base pair, gene sequences have been created using an overlapping primer PCR strategy. The nucleotide bias of these sequences was designed to have an E.coli codon usage bias.

For the first sequence, six oligonucleotides representing the first (5') 268 nucleotides of the native sequence for botulinum toxin type B were synthesised. For the second sequence 23 oligonucleotides representing internal sequence nucleotides 691-1641 of the native sequence for botulinum toxin type B were synthesised. The oligonucleotides ranged from 57-73 nucleotides in length. Overlapping regions, 17-20 nucleotides, were designed to give melting temperatures in the range 52-56°C. In addition, terminal restriction endonuclease sites of the synthetic products were constructed to facilitate insertion of these products into the exact corresponding region of the native sequence. The 268 bp 5' synthetic sequence has been incorporated into the gene shown in SEQ ID NO: 21 in place of the original first 268 bases (and is shown in SEQ ID NO: 27). Similarly the sequence could be inserted into other genes of the examples.

Another variant sequence equivalent to nucleotides 691 to 1641 of SEQ ID NO: 21

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- 25 , and employing non-native codon usage whilst coding for a native polypeptide sequence, has been constructed using the internal synthetic sequence. This sequence (SEQ ID NO: 28) can be incorporated, alone or in combination with other variant sequences, in place of the equivalent coding sequence in any of the genes of the example.

Example 3

An exemplification of the utility of this invention is as a non-toxic and effective immunogen. The non-toxic nature of the recombinant, single chain material was demonstrated by intraperitoneal administration in mice of GST-2LH423/A. The polypeptide was prepared and purified as described above. The amount of immunoreactive material in the final preparation was determined by enzyme linked immunosorbent assay (ELISA) using a monoclonal antibody (BA 11) reactive against a conformation dependent epitope on the native LHN/A. The recombinant material was serially diluted in phosphate buffered saline (PBS; NaCI 8 g/l, KCI 0.2 g/l, Na2HPO4 1.15 g/l, KH2PO4 0.2 g/l, pH 7.4) and 0.5 ml volumes injected into 3 groups of 4 mice such that each group of mice received 10, 5 and 1 micrograms of material respectively. Mice were observed for 4 days and no deaths were seen.

For immunisation, 20 pg of GST-2LH423/A in a 1.0 ml volume of water-in-oil emulsion (1:1 vokvol) using Freund's complete (primary injections only) or Freund's incomplete adjuvant was administered into guinea pigs via two sub-cutaneous dorsal injections. Three injections at 10 day intervals were given (day 1, day 10 and day 20) and antiserum collected on day 30. The antisera were shown by ELISA to be immunoreactive against native botulinum neurotoxin type A and to its derivative LHN/A. Antisera which were botulinum neurotoxin reactive at a dilution of 1:2000 were used for evaluation of neutralising efficacy in mice. For neutralisation assays 0.1 ml of antiserum was diluted into 2.5 ml of gelatine phosphate buffer (GPB; Na2HPO4 anhydrous 10 g/l, gelatin (Difco) 2 g/l, pH 6.56.6) containing a dilution range from 0.5 pg (5X1 O’6 g) to 5 picograms (5X1012 g). Aliquots of 0.5 ml were injected into mice intraperitoneally and deaths recorded

WO 98/07864 PCT/GB97/02273

- 26 over a 4 day period. The results are shown in Table 1 and Table 2. It can clearly be seen that 0.5 ml of 1:40 diluted anti- GST-2LH423/A antiserum can protect mice against intraperitoneal challenge with botulinum neurotoxin in the range 5 pg - 50 ng (1 - 10,000 mouse LD50; 1 mouse LD50 = 5 pg).

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- 27 TABLE 1 .

Neutralisation of botulinum neurotoxin in mice by guinea pig anti-GST-2LH423/A antiserum.

Botulinum Toxin/mouse

<td> Survivors</td><td> Ο.5μ9</td><td> Ο.ΟΟδμς</td><td> O.OOOSpg</td><td> O.5ng</td><td> 0.005ng</td><td> 5pg</td><td> Control</td>

<td> On Day</td><td></td><td></td><td></td><td></td><td></td><td></td><td> (no toxin)</td>

TABLE 2. Neutralisation of botulinum neurotoxin in mice by non-immune guinea pig antiserum.

Botulinum Toxin/mouse

<td> Survivors</td><td> 0.5pg</td><td> 0.005/jg</td><td> 0.0005pg</td><td> 0.5ng</td><td> 0.005ng</td><td> 5pg</td><td> Control</td>

<td> On Day</td><td></td><td></td><td></td><td></td><td></td><td></td><td> (no toxin)</td>

Example 4

Expression of recombinant LH107/B in E. coli.

As an exemplification of the expression of a nucleic acid coding for a LHN of a clostridial neurotoxin of a serotype other than botulinum neurotoxin type A, the nucleic acid sequence (SEQ ID NO; 23) coding for the polypeptide LH107/B (SEQ ID

WO 98/07864 PCT/GB97/02273

- 28 NO: 24) was inserted into the commercially available plasmid pET28a (Novogen, Madison, Wl, USA). The nucleic acid was expressed in E. coli BL21 (DE3) (New England BioLabs, Beverley, MA, USA) as a fusion protein with a N-terminal T7 fusion peptide, under IPTG induction at 1 mM for 90 minutes at 37°C. Cultures were harvested and recombinant protein extracted as described previously for LH423/A.

Recombinant protein was recovered and purified from bacterial paste lysates by immunoaffinity adsorption to an immobilised anti-T7 peptide monoclonal antibody using aT7 tag purification kit (New England bioLabs, Beverley, MA, USA). Purified recombinant protein was analysed by gradient (4-20%) denaturing SDSpolyacrylamide gel electrophoresis (Novex, San Diego, CA, USA) and western blotting using polyclonal anti-botulinum neurotoxin type antiserum or anti-T7 antiserum. Western blotting reagents were from Novex, immunostained proteins were visualised using the Enhanced Chemi-Luminescence system (ECL) from Amersham. The expression of an anti-T7 antibody and anti-botulinum neurotoxin type B antiserum reactive recombinant product is demonstrated in Figure 13.

The recombinant product was soluble and retained that part of the light chain responsible for endopeptidase activity.

The invention thus provides recombinant polypeptides useful inter alia as immunogens, enzyme standards and components for synthesis of molecules as described in WO-A-94/21300.

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- 29 SEQUENCE LISTING (1) GENERAL INFORMATION:

(i) APPLICANT:

(A) NAME: MICROBIOLOGICAL RESEARCH AUTHORITY (B) STREET: Centre For Applied Microbiology And Research,

Porton Down (C) CITY: Salisbury (D) STATE: Wiltshire (E) COUNTRY: UK (F) POSTAL CODE (ZIP): SP4 OJG (A) NAME: THE SPEYWOOD LABORATORY LIMITED (B) STREET: 14 Kensington Square (C) CITY: London (E) COUNTRY: UK (F) POSTAL CODE (ZIP): W8 5HH (A) NAME: FOSTER; Keith Alan (B) STREET: Centre For Applied Microbiology And Research,

Porton Down (C) CITY: Salisbury (D) STATE: Wiltshire (E) COUNTRY: UK (F) POSTAL CODE (ZIP): SP4 OJG (A) NAME: QUINN; Conrad Padraig (B) STREET: Centre For Applied Microbiology And Research,

Porton Down (C) CITY: Salisbury (D) STATE: Wiltshire (E) COUNTRY: UK (F) POSTAL CODE (ZIP): SP4 OJG (A) NAME: SHONE; Clifford Charles (B) STREET: Centre For Applied Microbiology And Research,

Porton Down (C) CITY: Salisbury (D) STATE: Wiltshire (E) . COUNTRY: UK (F) POSTAL CODE (ZIP): SP4 OJG (ii) TITLE OF INVENTION: Recombinant Toxin Fragments (iii) NUMBER OF SEQUENCES: 28 (iv) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.30 (EPO) (2) INFORMATION FOR SEQ ID NO: 1:

(i) SEQUENCE CHARACTERISTICS :

(A) LENGTH: 2616 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic)

- 30 (ix) FEATURE:

(A) NAME/KEY : CDS (B) LOCATION:1. .2616 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:

ATG CAG TTC GTG AAC AAG CAG TTC AAC TAT AAG GAC CCT GTA AAC GGT48

Met Gin Phe Val Asn Lys Gin Phe Asn Tyr Lys Asp Pro Val AsnGly

10is

GTT GAC ATT GCC TAC ATC AAA ATT CCA AAC GCC GGC CAG ATG CAG CCG96

Val Asp He Ala Tyr He Lys He Pro Asn Ala Gly Gin Met GinPro

2530

GTG AAG GCT TTC AAG ATT CAT AAC AAA ATC TGG GTT ATT CCG GAA CGC144

Val Lys Ala Phe Lys lie His Asn Lys He Trp Val He Pro GluArg

4045

GAT ACA TTT ACG AAC CCG GAA GAA GGA GAC TTG AAC CCG CCG CCG GAA192

Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro ProGlu

5560

GCA AAG CAG GTG CCA GTT TCA TAC TAC GAT TCA ACC TAT CTG AGC ACA24 0

Ala Lys Gin Val Pro Val Ser Tyr Tyr Asp Ser Thr Tyr Leu SerThr

70 75qq

GAC AAC GAG AAG GAT AAC TAC CTG AAG GGA GTG ACC AAA TTA TTC GAG288

Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Val Thr Lys Leu PheGlu

9095

CGT ATT TAT TCC ACT GAC CTG GGC CGT ATG CTG CTG ACC TCA ATC GTC336

Arg lie Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser lieVal

100 105no

CGC GGA ATC CCA TTT TGG GGT GGC AGT ACC ATT GAC ACG GAG TTG AAG384

Arg Gly He Pro Phe Trp Gly Gly Ser Thr lie Asp Thr Glu LeuLys

115 120125

GTT ATT GAC ACT AAC TGC ATT AAC GTG ATC CAA CCA GAC GGT AGC TAC432

Val lie Asp Thr Asn Cys lie Asn Val lie Gin Pro Asp Gly SerTyr

130 135140

AGA TCT GAA GAA CTT AAC CTC GTA ATC ATC GGG CCC TCC GCG GAC ATT480

Arg Ser Glu Glu Leu Asn Leu Val lie lie Gly Pro Ser Ala AspHe

145 150 155160

ATC CAG TTT GAG TGC AAG AGC TTT GGC CAC GAA GTG TTG AAC CTG ACG528 lie Gin Phe Glu Cys Lys Ser Phe Gly His Glu Val Leu Asn LeuThr

165 170175

CGT AAC GGT TAC GGC TCT ACT CAG TAC ATT CGT TTC AGC CCA GAC TTC576

Arg Asn Gly Tyr Gly Ser Thr Gin Tyr lie Arg Phe Ser Pro AspPhe

180 185190

ACG TTC GGT TTC GAG GAG AGC CTG GAG GTT GAT ACC AAC CCG CTG TTG624

Thr Phe Gly Phe Glu Glu Ser Leu Glu Val Asp Thr Asn Pro LeuLeu

195 200205

GGT GCA GGC AAG TTC GCA ACT GAT CCA GCG GTG ACC CTG GCA CAC GAG672

Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Val Thr Leu Ala HisGlu

210 215220

CTG ATC CAC GCC GGT CAT CGT CTG TAT GGC ATT GCG ATT AAC CCG AAC720

Leu lie His Ala Gly His Arg Leu Tyr Gly He Ala He Asn ProAsn

225 230 235240

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- 31 CGC GTG TTC AAG GTT AAC ACC AAC GCC TAC TAC GAG ATG AGT GGT TTA768

Arg Vai Phe Lys Vai Asn Thr Asn Ala Tyr Tyr Glu Met Ser Gly Leu 245 250255

GAA GTA AGC TTC GAG GAA CTG CGC ACG TTC GGT GGC CAT GAT GCG AAG816

Glu Vai Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp AlaLys

260 265270

TTT ATC GAC AGC TTG CAG GAG AAC GAG TTC CGT CTG TAC TAC TAC AAC864

Phe He Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr TyrAsn

275 280285

AAG TTT AAA GAT ATT GCA AGT ACA CTG AAC AAG GCT AAG TCC ATT GTG912

Lys Phe Lys Asp He Ala Ser Thr Leu Asn Lys Ala Lys Ser lieVai

290 295300

GGT ACC ACT GCT TCA TTA CAG TAT ATG AAA AAT GTT TTT AAA GAG AAA960

Gly Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn Vai Phe Lys GluLys

305 310 315220

TAT CTC CTA TCT GAA GAT ACA TCT GGA AAA TTT TCG GTA GAT. AAA TTA1008

Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys’ Phe Ser Vai Asp LysLeu

325 330335

AAA TTT GAT AAG TTA TAC AAA ATG TTA ACA GAG ATT TAC ACA GAG GAT1056

Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu lie Tyr Thr GluAsp

340 345350

AAT TTT GTT AAG TTT TTT AAA GTA CTT AAC AGA AAA ACA TAT TTG AAT1104

Asn Phe Vai Lys Phe Phe Lys Vai Leu Asn Arg Lys Thr Tyr LeuAsn

355 360365

TTT GAT AAA GCC GTA TTT AAG ATA AAT ATA GTA CCT AAG GTA AAT TAC1152

Phe Asp Lys Ala Vai Phe Lys He Asn He Vai Pro Lys Vai AsnTyr

370 375380

ACA ATA TAT GAT GGA TTT AAT TTA AGA AAT ACA AAT TTA GCA GCA AAC1200

Thr lie Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala AlaAsn

385 390 395400

TTT AAT GGT CAA AAT ACA GAA ATT AAT AAT ATG AAT TTT ACT AAA CTA124 8

Phe Asn Gly Gin Asn Thr Glu He Asn Asn Met Asn Phe Thr LysLeu

405 410415

AAA AAT TTT ACT GGA TTG TTT GAA TTT TAT AAG TTG CTA TGT GTA AGA1296

Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys VaiArg

420 425430

GGG ATA ATA ACT TCT AAA ACT AAA TCA TTA GAT AAA GGA TAC AAT AAG1344

Gly He He Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr AsnLys

435 440445

GCA TTA AAT GAT TTA TGT ATC AAA GTT AAT AAT TGG GAC TTG TTT TTT1392

Ala Leu Asn Asp Leu Cys lie Lys Vai Asn Asn Trp Asp Leu PhePhe

450 455460

AGT CCT TCA GAA GAT AAT TTT ACT AAT GAT CTA AAT AAA GGA GAA GAA1440

Ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn Lys Gly GluGlu

465 470 475480

ATT ACA TCT GAT ACT AAT ATA GAA GCA GCA GAA GAA AAT ATT AGT TTA14 88 lie Thr Ser Asp Thr Asn He Glu Ala Ala Glu Glu Asn He SerLeu

485 490495

GAT TTA ATA CAA CAA TAT TAT TTA ACC TTT AAT TTT GAT AAT GAA CCT1536

Asp Leu lie Gin Gin Tyr Tyr Leu Thr Phe Asn Phe Asp Asn GluPro

500 505510

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- 33 AAT AAA TTT TTG AAT CAA TGC TCT GTT TCA TAT TTA ATG AAT TCT ATG2400

Asn Lys Phe Leu Asn Gin Cys Ser Vai Ser Tyr Leu Met Asn SerMet

785 790 795800

ATC CCT TAT GGT GTT AAA CGG TTA GAA GAT TTT GAT GCT AGT CTT AAA244 8 lie Pro Tyr Gly Vai Lys Arg Leu Glu Asp Phe Asp Ala Ser LeuLys

805 810815

GAT GCA TTA TTA AAG TAT ATA TAT GAT AAT AGA GGA ACT TTA ATT GGT24 96

Asp Ala Leu Leu Lys Tyr He Tyr Asp Asn Arg Gly Thr Leu HeGly

820 825830

CAA GTA GAT AGA TTA AAA GAT AAA GTT AAT AAT ACA CTT AGT ACA GAT2544

Gin Vai Asp Arg Leu Lys Asp Lys Vai Asn Asn Thr Leu Ser ThrAsp

835 840845

ATA CCT TTT CAG CTT TCC AAA TAC GTA GAT AAT CAA AGA TTA TTA TCT2592 lie Pro Phe Gin Leu Ser Lys Tyr Vai Asp Asn Gin Arg Leu LeuSer

850 855860

ACA TTT ACT GAA TAT ATT AAG TAA2616

Thr Phe Thr Glu Tyr lie Lys * 865870 (2) INFORMATION FOR SEQ ID NO: 2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 872 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:

Met Gin Phe Vai Asn Lys Gin Phe Asn Tyr Lys Asp Pro Vai Asn Gly 15 1015

Vai Asp lie Ala Tyr lie Lys lie Pro Asn Ala Gly Gin Met Gin Pro 20 2530

Vai Lys Ala Phe Lys lie His Asn Lys He Trp Vai lie Pro Glu Arg 35 4045

Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 5560

Ala Lys Gin Vai Pro Vai Ser Tyr Tyr Asp Ser Thr Tyr Leu Ser Thr

70 7580

Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Vai Thr Lys Leu Phe Glu 85 9095

Arg lie Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser lie Vai 100 105HO

Arg Gly lie Pro Phe Trp Gly Gly Ser Thr He Asp Thr Glu Leu Lys 115 120125

Vai lie Asp Thr Asn Cys lie Asn Vai lie Gin Pro Asp Gly Ser Tyr 130 135140

Arg Ser Glu Glu Leu Asn Leu Vai lie lie Gly Pro Ser Ala Asp lie

145 150 155160 lie Gin Phe Glu Cys Lys Ser Phe Gly His Glu Vai Leu Asn Leu Thr

165 170175

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Arg Asn

Thr Phe

Gly Ala

210

Leu lie 225

Arg Vai

Glu Vai

Phe He

Lys Phe

290

Gly Thr 305

Tyr Leu

Lys Phe

Asn Phe

Phe Asp

370

Thr He 385

Phe Asn

Lys Asn

Gly He

Ala Leu

450

Ser Pro 465

He Thr

Asp Leu

Gly Tyr

180

Gly Phe 195

Gly Lys

His Ala

Phe Lys

Ser Phe

260

Asp Ser 275

Lys Asp

Thr Ala

Leu Ser

Asp Lys

340

Vai Lys 355

Lys Ala

Tyr Asp

Gly Gin

Phe Thr

420

He Thr 435

Asn Asp

Ser Glu

Ser Asp

He Gin

500

Gly Ser

Glu Glu

Phe Ala

Gly His

230

Vai Asn 245

Glu Glu

Leu Gin lie Ala

Ser Leu

310

Glu Asp 325

Leu Tyr

Phe Phe

Vai Phe

Gly Phe

390

Asn Thr 405

Gly Leu

Ser Lys

Leu Cys

Asp Asn

470

Thr Asn 485

Gin Tyr

Thr Gin

Ser Leu

200

Thr Asp 215

Arg Leu

Thr Asn

Leu Arg

Glu Asn

280

Ser Thr 295

Gin Tyr

Thr Ser

Tyr He 185

Glu Vai

Pro Ala

Tyr Gly

Ala Tyr

250

Thr Phe 265

Glu Phe

Leu Asn

Met Lys

Lys Met

Lys Vai

360

Lys lie

375

Asn Leu

Glu He

Phe Glu

Thr Lys

440

He Lys

455

Phe Thr

He Glu

Tyr Leu

Gly Lys

330

Leu Thr 345

Leu Asn

Asn lie

Arg Asn

Asn ASn

410

Phe Tyr

425

Ser Leu

Vai Asn

Asn Asp

Ala Ala

490

Thr Phe 505

- 34 Arg Phe

Asp Thr

Vai Thr

220

He Ala

235

Tyr Glu

Gly Gly

Arg Leu

Lys Ala

300

Asn Vai 315

Phe Ser

Glu He

Arg Lys

Vai Pro

380

Thr Asn 395

Met Asn

Lys Leu

Asp Lys

Asn Trp

460

Leu Asn 475

Glu Glu

Asn Phe

Ser Pro

190

Asn Pro 205

Leu Ala

He Asn

Met Ser

His Asp

270

Tyr Tyr 285

Lys Ser

Phe Lys

Vai Asp

Tyr Thr

350

Thr Tyr 365

Lys Vai

Leu Ala

Phe Thr

Leu Cys

430

Gly Tyr

445

Asp Leu

Lys Gly

Asn He

Asp Asn

510

Asp Phe

Leu Leu

His Glu

Pro Asn

240

Gly Leu 255

Ala Lys

Tyr Asn

He Vai

Glu Lys

320

Lys Leu 335

Glu Asp

Leu Asn

Asn Tyr

Ala Asn

400

Lys Leu

415

Vai Arg

Asn Lys

Phe Phe

Glu Glu

480

Ser Leu 495

G1U Pro

Glu Asn lie Ser He Glu Asn Leu Ser Ser Asp lie He Gly Gin Leu

515 520 525

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865 870 (2) INFORMATION FOR SEQ ID NO: 3:

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PCT/GB97/02273

- 36 (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2685 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE:

(A) NAME/KEY: CDS (B) LOCATION:!..2685 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:

WO 98/07864

PCT/GB97/02273

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PCT/GB97/02273

740 745 750

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PCT/GB97/02273

- 39 AAA AAA ATG AAA GAA GCT TTA GAA AAT CAA GCA GAA GCA ACA AAG GCT

Lys Lys Met Lys Glu Ala Leu Glu Asn Gin Ala Glu Ala Thr Lys Ala

755 76076S

ATA ATA AAC TAT CAG TAT AAT CAA TAT ACT GAG GAA GAG AAA AATAAT

He lie Asn Tyr Gin Tyr Asn Gin Tyr Thr Glu Glu Glu Lys AsnAsn

770 775780

ATT AAT TTT AAT ATT GAT GAT TTA AGT TCG AAA CTT AAT GAG TCTATA

He Asn Phe Asn lie Asp Asp Leu Ser Ser Lys Leu Asn Glu SerHe 785 790 795800

AAT AAA GCT ATG ATT AAT ATA AAT AAA TTT TTG AAT CAA TGC TCTGTT

Asn Lys Ala Met lie Asn lie Asn Lys Phe Leu Asn Gin Cys SerVai

805 810815

TCA TAT TTA ATG AAT TCT ATG ATC CCT TAT GGT GTT AAA CGG TTAGAA

Ser Tyr Leu Met Asn Ser Met He Pro Tyr Gly Vai Lys Arg LeuGlu

820 825830

GAT TTT GAT GCT AGT CTT AAA GAT GCA TTA TTA AAG TAT ATA TATGAT

Asp Phe Asp Ala Ser Leu Lys Asp Ala Leu Leu Lvs Tyr lie TyrAsp

835 840 '845

AAT AGA GGA ACT TTA ATT GGT CAA GTA GAT AGA TTA AAA GAT AAAGTT

Asn Arg Gly Thr Leu lie Gly Gin Vai Asp Arg Leu Lys Asp LysVai

850 855860

AAT AAT ACA CTT AGT ACA GAT ATA CCT TTT CAG CTT TCC AAA TACGTA

Asn Asn Thr Leu Ser Thr Asp He Pro Phe Gin Leu Ser Lys TyrVai

865 870 875880

GAT AAT CAA AGA TTA TTA TCT ACA TTT ACT GAA TAT ATT AAGTAA

Asp Asn Gin Arg Leu Leu Ser Thr Phe Thr Glu Tyr He Lys*

885 890895

2304

2352

2400

2448

2496

2544

2592

2640

2685 (2) INFORMATION FOR SEQ ID NO: 4:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 895 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:

Asp Ser Thr Tyr Leu Ser Thr Asp Asn Glu Lys Asp Asn Tyr Leu Lys

100 105 110

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<td> (2)</td><td colspan="3"> INFORMATION</td><td> FOR</td><td> SEQ</td><td colspan="2"> ID NO: 5</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

(i) SEQUENCE CHARACTERISTICS:

<td rowspan="3"></td><td colspan="3"> (A) LENGTH: 2622 base pairs</td><td rowspan="3"></td><td rowspan="3"></td><td rowspan="3"></td><td rowspan="3"></td><td rowspan="3"></td><td rowspan="3"></td><td rowspan="3"></td>

<td colspan="2"> (B) TYPE: nucleic acid (C) STRANDEDNESS: double</td><td rowspan="2"> c)</td>

<td> (D) TOPOLOGY: (ii) MOLECULE TYPE: (ix) FEATURE: (A) NAME/KEY: (B) LOCATION:</td><td> linear DNA (genomi CDS 1. .2622</td>

<td></td><td colspan="2"> (xi) SEQUENCE DESCRIPTION: SEQ</td><td colspan="2"> ID NO: 5</td><td></td><td></td><td></td><td></td><td></td><td></td>

WO 98/07864

PCT/GB97/02273

AGC TAC AGA TCT GAA GAA CTT AAC CTC

Ser Tyr Arg Ser Glu Glu Leu Asn Leu

145 150

GAC ATT ATC CAG TTT GAG TGC AAG AGC

Asp He He Gin Phe Glu Cys Lys Ser

165

CTG ACG CGT AAC GGT TAC GGC TCT ACT

Leu Thr Arg Asn Gly Tyr Gly Ser Thr

180185

GAC TTC ACG TTC GGT TTC GAG GAGAGC

Asp Phe Thr Phe Gly Phe Glu GluSer

195200

CTG TTG GGT GCA GGC AAG TTC GCAACT

Leu Leu Gly Ala Gly Lys Phe AlaThr

210215

CAC GAG CTG ATC CAC GCC GGT CATCGT

His Glu Leu He His Ala Gly HisArg

225230

CCG AAC CGC GTG TTC AAG GTT AACACC

Pro Asn Arg Vai Phe Lys Vai Asn Thr

245

GGT TTA GAA GTA AGC TTC GAG GAA CTG

Gly Leu Glu Vai Ser Phe Glu Glu Leu

260265

GCG AAG TTT ATC GAC AGC TTG CAGGAG

Ala Lys Phe lie Asp Ser Leu GinGlu

275280

TAC AAC AAG TTT AAA GAT ATT GCAAGT

Tyr Asn Lys Phe Lys Asp He AlaSer

290295

ATT GTG GGT ACC ACT GCT TCA TTACAG lie Vai Gly Thr Thr Ala Ser LeuGin

305310

GAG AAA TAT CTC CTA TCT GAA GAT ACA

Glu Lys Tyr Leu Leu Ser Glu Asp Thr

325

AAA TTA AAA TTT GAT AAG TTA TAC AAA

Lys Leu Lys Phe Asp Lys Leu TyrLys

340345

GAG GAT AAT TTT GTT AAG TTT TTTAAA

Glu Asp Asn Phe Vai Lys Phe PheLys

355360

TTG AAT TTT GAT AAA GCC GTA TTTAAG

Leu Asn Phe Asp Lys Ala Vai PheLys

370375

AAT TAC ACA ATA TAT GAT GGA TTTAAT

Asn Tyr Thr He Tyr Asp Gly PheAsn

385390

GCA AAC TTT AAT GGT CAA AAT ACA GAA

Ala Asn Phe Asn Gly Gin Asn Thr Glu

405

- 43 GTA ATC ATC GGG CCC TCC GCG4 80

Vai lie He Gly Pro Ser Ala 155160

TTT GGC CAC GAA GTG TTG AAC52 8

Phe Gly His Glu Vai Leu Asn 170175

CAG TAC ATT CGT TTC AGC CCA576

Gin Tyr He Arg Phe Ser Pro

190

CTG GAG GTT GAT ACC AAC CCG624

Leu Glu Vai Asp Thr Asn Pro

205

GAT CCA GCG GTG ACC CTG GCA672

Asp Pro Ala Vai Thr Leu Ala 220

CTG TAT GGC ATT GCG ATT AAC720

Leu Tyr Gly He Ala He Asn 235240

AAC GCC TAC TAC GAG ATG AGT76 8

Asn Ala Tyr Tyr Glu Met Ser 250255

CGC ACG TTC GGT GGC CAT GAT816

Arg Thr Phe Gly Gly His Asp

270

AAC GAG TTC CGT CTG TAC TAC864

Asn Glu Phe Arg Leu Tyr Tyr

285

ACA CTG AAC AAG GCT AAG TCC912

Thr Leu Asn Lys Ala Lys Ser 300

TAT ATG AAA AAT GTT TTT AAA960

Tyr Met Lys Asn Vai Phe Lys

315320

TCT GGA AAA TTT TCG GTA GAT 100 8

Ser Gly Lys Phe Ser Vai Asp 330335

ATG TTA ACA GAG ATT TAC ACA 1056

Met Leu Thr Glu He Tyr Thr

350

GTA CTT AAC AGA AAA ACA TAT 1104

Vai Leu Asn Arg Lys Thr Tyr

365

ATA AAT ATA GTA CCT AAG GTA 1152

He Asn lie Vai Pro Lys Vai 380

TTA AGA AAT ACA AAT TTA GCA 1200

Leu Arg Asn Thr Asn Leu Ala

395 400

ATT AAT AAT ATG AAT TTT ACT 1248 He Asn Asn Met Asn Phe Thr 410 415

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- 44 AAA CTA AAA AAT TTT ACT GGA TTG TTT GAA TTT TAT AAG TTG CTA TGT1296

Lys Leu Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu LeuCys

420 425430

GTA AGA GGG ATA ATA ACT TCT AAA ACT AAA TCA TTA GAT AAA GGA TAC1344

Val Arg Gly He lie Thr Ser Lys Thr Lys Ser Leu Asp Lys GlyTyr

435 440445

AAT AAG GCA TTA AAT GAT TTA TGT ATC AAA GTT AAT AAT TGG GAC TTG1392

Asn Lys Ala Leu Asn Asp Leu Cys lie Lys Val Asn Asn Trp AspLeu

450 455460

TTT TTT AGT CCT TCA GAA GAT AAT TTT ACT AAT GAT CTA AAT AAA GGA1440

Phe Phe Ser Pro Ser Glu Asp Asn Phe Thr Asn Asp Leu Asn LysGly

465 470 475480

GAA GAA ATT ACA TCT GAT ACT AAT ATA GAA GCA GCA GAA GAA AAT ATT1488

Glu Glu lie Thr Ser Asp Thr Asn lie Glu Ala Ala Glu Glu AsnHe

485 490495

AGT TTA GAT TTA ATA CAA CAA TAT TAT TTA ACC TTT AAT TTT GAT AAT1536

Ser Leu Asp Leu He Gin Gin Tyr Tyr Leu Thr Phe Asn Phe AspAsn

500 505510

GAA CCT GAA AAT ATT TCA ATA GAA AAT CTT TCA AGT GAC ATT ATA GGC1584

Glu Pro Glu Asn lie Ser lie Glu Asn Leu Ser Ser Asp He lieGly

515 520525

CAA TTA GAA CTT ATG CCT AAT ATA GAA AGA TTT CCT AAT GGA AAA AAG163 2

Gin Leu Glu Leu Met Pro Asn lie Glu Arg Phe Pro Asn Gly LysLys

530 535540

TAT GAG TTA GAT AAA TAT ACT ATG TTC CAT TAT CTT CGT GCT CAA GAA1680

Tyr Glu Leu Asp Lys Tyr Thr Met Phe His Tyr Leu Arg Ala GinGlu

545 550 555560

TTT GAA CAT GGT AAA TCT AGG ATT GCT TTA ACA AAT TCT GTT AAC GAA172 8

Phe Glu His Gly Lys Ser Arg lie Ala Leu Thr Asn Ser Val AsnGlu

565 570575

GCA TTA TTA AAT CCT AGT CGT GTT TAT ACA TTT TTT TCT TCA GAC TAT1776

Ala Leu Leu Asn Pro Ser Arg Val Tyr Thr Phe Phe Ser Ser AspTyr

580 585590

GTA AAG AAA GTT AAT AAA GCT ACG GAG GCA GCT ATG TTT TTA GGC TGG1824

Val Lys Lys Val Asn Lys Ala Thr Glu Ala Ala Met Phe Leu GlyTrp

595 600605

GTA GAA CAA TTA GTA TAT GAT TTT ACC GAT GAA ACT AGC GAA GTA AGT1872

Val Glu Gin Leu Val Tyr Asp Phe Thr Asp Glu Thr Ser Glu ValSer

610 615620

ACT ACG GAT AAA ATT GCG GAT ATA ACT ATA ATT ATT CCA TAT ATA GGA1920

Thr Thr Asp Lys He Ala Asp lie Thr lie He lie Pro Tyr lieGly

625 630 635640

CCT GCT TTA AAT ATA GGT AAT ATG TTA TAT AAA GAT GAT TTT GTA GGT1968

Pro Ala Leu Asn He Gly Asn Met Leu Tyr Lys Asp Asp Phe ValGly

645 650655

GCT TTA ATA TTT TCA GGA GCT GTT ATT CTG TTA GAA TTT ATA CCA GAG2016

Ala Leu lie Phe Ser Gly Ala Val lie Leu Leu Glu Phe lie ProGlu

660 665670

ATT GCA ATA CCT GTA TTA GGT ACT TTT GCA CTT GTA TCA TAT ATT GCG2064 lie Ala lie Pro Val Leu Gly Thr Phe Ala Leu Val Ser Tyr HeAla

675 680685

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865 870 (2) INFORMATION FOR SEQ ID NO: 6:

(i) SEQUENCE CHARACTERISTICS :

(A) LENGTH: 874 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:

WO 98/07864 . c PCT/GB97/02273

Asn Tyr Thr He Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala 385 390 395 400

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PCT/GB97/O2273

740 745 750

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<td colspan="2"> Thr Asp lie Pro Phe Gin Leu Ser Lys Tyr Vai Asp Asn Gin Arg Leu 850 855 860 Leu Ser Thr Phe Thr Glu Tyr lie Lys * 865 870 (2) INFORMATION FOR SEQ ID NO: 7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2613 base pairs - (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:!..2613 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7:</td>

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- 49 AGA ATT TAT TCA ACT GAT CTT GGA AGA ATG TTG TTA ACA TCA ATA GTA

Arg He Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser lieVai

100 105HO

AGG GGA ATA CCA TTT TGG GGT GGA AGT ACA ATA GAT ACA GAA TTAAAA

Arg Gly He Pro Phe Trp Gly Gly Ser Thr lie Asp Thr Glu LeuLvs 115 120125

GTT ATT GAT ACT AAT TGT ATT AAT GTG ATA CAA CCA GAT GGT AGTTAT

Vai He Asp Thr Asn Cys lie Asn Vai He Gin Pro Asp Glv SerTvr

130 135 140

AGA TCA GAA GAA CTT AAT CTA GTA ATA ATA GGA CCC TCA GCT GAT ATT

Arg Ser Glu Glu Leu Asn Leu Vai He lie Gly Pro Ser Ala Asn He

145 150 155 2gQ

ATA CAG TTT GAA TGT AAA AGC TTT GGA CAT GAA GTT TTG AAT CTT ACG

He Gin Phe Glu Cys Lys Ser Phe Gly His Glu Vai Leu Asn Leu Thr

165 170175

CGA AAT GGT TAT GGC TCT ACT CAA TAC ATT AGA TTT AGC CCA GATTTT

Arg Asn Gly Tyr Gly Ser Thr Gin Tyr He Arg Phe Ser Pro AspPhe

180 185j_90

ACA TTT GGT TTT GAG GAG TCA CTT GAA GTT GAT ACA AAT CCT CTTTTA

Thr Phe Gly Phe Glu Glu Ser Leu Glu Vai Asp Thr Asn Pro LeuLeu

195 200205

GGT GCA GGC AAA TTT GCT ACA GAT CCA GCA GTA ACA TTA GCA CATGAA

Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Vai Thr Leu Ala HisGlu

210 215220

CTT ATA CAT GCT GGA CAT AGA TTA TAT GGA ATA GCA ATT AAT CCAAAT

Leu He His Ala Gly His Arg Leu Tyr Gly He Ala He Asn ProAsn

225 230 235240

AGG GTT TTT AAA GTA AAT ACT AAT GCC TAT TAT GAA ATG AGT GGGTTA

Arg Vai Phe Lys Vai Asn Thr Asn Ala Tyr Tyr Glu Met Ser GlyLeu

245 250255

GAA GTA AGC TTT GAG GAA CTT AGA ACA TTT GGG GGA CAT GAT GCAAAG

Glu Vai Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp AlaLvs

260 265270

TTT ATA GAT AGT TTA CAG GAA AAC GAA TTT CGT CTA TAT TAT TATAAT

Phe lie Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr TyrAsn

275 280285

AAG TTT AAA GAT ATA GCA AGT ACA CTT AAT AAA GCT AAA TCA ATAGTA

Lys Phe Lys Asp He Ala Ser Thr Leu Asn Lys Ala Lys Ser HeVai

290 295300

GGT ACT ACT GCT TCA TTA CAG TAT ATG AAA AAT GTT TTT AAA GAGAAA

Gly Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn Vai Phe Lys GluLys

305 310 315

TAT CTC CTA TCT GAA GAT ACA TCT GGA AAA TTT TCG GTA GAT AAATTA

Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Vai Asp LysLeu

325 330335

AAA TTT GAT AAG TTA TAC AAA ATG TTA ACA GAG ATT TAC ACA GAGGAT

Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu He Tyr Thr GluAsp

340 345350

336

384

432

480

528

576

624

672

720

768

816

864

912

960

1008

1056

AAT TTT GTT AAG TTT TTT AAA GTA CTT AAC AGA AAA ACA TAT TTG AAT Asn Phe Vai Lys Phe Phe Lys Vai Leu Asn Arg Lys Thr Tyr Leu Asn 355 360 365

1104

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PCT/GB97/02273

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PCT/GB97/02273

- 5Ί -

865 870 (2) INFORMATION FOR SEQ ID NO: 8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 871 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear

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PCT/GB97/02273

- 52 (ii) MOLECULE TYPE: protein (Xi) SEQUENCE DESCRIPTION: SEQ ID NO : 8:

Met Pro Phe Vai Asn Lys Gin Phe Asn Tyr Lys Asp Pro Vai AsnGlv 1 5 1015

Vai Asp He Ala Tyr lie Lys lie Pro Asn Ala Gly Gin Met GinPro

2530

Vai Lys Ala Phe Lys lie His Asn Lys lie Trp Vai He Pro Glu Arq 35 4045

Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro Pro Glu 50 5560

Ala Lys Gin Vai Pro Vai Ser Tyr Tyr Asp Ser Thr Tyr Leu SerThr

70 7580

Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Vai Thr Lys Leu PheGlu 85 9095

Arg He Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser lie Vai 100 105no

Arg Gly He Pro Phe Trp Gly Gly Ser Thr He Asp Thr Glu Leu Lys 115 120125

Vai He Asp Thr Asn Cys lie Asn Vai He Gin Pro Asp Gly Ser Tyr 130 135140

Arg Ser Glu Glu Leu Asn Leu Vai lie He Gly Pro Ser Ala Asplie

145 150 155150 lie Gin Phe Glu Cys Lys Ser Phe Gly His Glu Vai Leu Asn LeuThr

165 170175

Arg Asn Gly Tyr Gly Ser Thr Gin Tyr lie Arg Phe Ser Pro Asp Phe 180 185190

Thr Phe Gly Phe Glu Glu Ser Leu Glu Vai Asp Thr Asn Pro Leu Leu 195 200205

Gly Ala Gly Lys Phe Ala Thr Asp Pro Ala Vai Thr Leu Ala His Glu 210 - 215220

Leu He His Ala Gly His Arg Leu Tyr Gly He Ala He Asn ProAsn

225 230 235240

Arg Vai Phe Lys Vai Asn Thr Asn Ala Tyr Tyr Glu Met Ser GlyLeu

245 250255

Glu Vai Ser Phe Glu Glu Leu Arg Thr Phe Gly Gly His Asp Ala Lys 260 265270

Phe lie Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn 275 280285

Lys Phe Lys Asp He Ala Ser Thr Leu Asn Lys Ala Lys Ser He Vai 290 295300

Gly Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn Vai Phe Lys GluLys

305 310 315320

Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Vai Asp LysLeu

325 330335

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Lys Phe Asp Lys Leu Tyr Lys Met 340

Asn Phe Vai Lys Phe Phe Lys Vai

355360

Phe Asp Lys Ala Vai Phe Lys He 370375

Thr lie Tyr Asp Gly Phe Asn Leu

385390

Phe Asn Gly Gin Asn Thr Glu He 405

Lys Asn Phe Thr Gly Leu Phe Glu 420

Gly He lie Thr Ser Lys Thr Lys

435440

Ala Leu Asn Asp Leu Cys He Lys 450455

Ser Pro Ser Glu Asp Asn Phe Thr

465470 lie Thr Ser Asp Thr Asn He Glu

485

Asp Leu lie Gin Gin Tyr Tyr Leu 500

Glu Asn lie Ser He Glu Asn Leu

515520

Glu Leu Met Pro Asn lie Glu Arg 530535

Leu Asp Lys Tyr Thr Met Phe His

545550

His Gly Lys Ser Arg lie Ala Leu 565

Leu Asn Pro Ser Arg Vai Tyr Thr 580

Lys Vai Asn Lys Ala Thr Glu Ala

595600

Gin Leu Vai Tyr Asp Phe Thr Asp 610615

Asp Lys lie Ala Asp He Thr He

625630

Leu Asn He Gly Asn Met Leu Tyr 645 lie Phe Ser Gly Ala Vai lie Leu 660

He Pro Vai Leu Gly Thr Phe Ala

675 680

- 53 Leu Thr Glu lie Tyr Thr Glu Asp

345 35Q

Leu Asn Arg Lys Thr Tyr Leu Asn 365

Asn lie Vai Pro Lys Vai Asn Tyr 380

Arg Asn Thr Asn Leu Ala Ala Asn 3954

Asn Asn Met Asn Phe Thr Lvs Leu 410415

Phe Tyr Lys Leu Leu Cys Vai Arq

42543o

Ser Leu Asp Lys Gly Tyr Asn Lys 445

Vai Asn Asn Trp Asp Leu Phe Phe 460

Asn Asp Leu Asn Lys Gly Glu Glu

475400

Ala Ala Glu Glu Asn He Ser Leu 490495

Thr Phe Asn Phe Asp Asn Glu Pro

5055io

Ser Ser Asp lie lie Gly Gin Leu 525

Phe Pro Asn Gly Lys Lys Tyr Glu 540

Tyr Leu Arg Ala Gin Glu Phe Glu

555560

Thr Asn Ser Vai Asn Glu Ala Leu 570575

Phe Phe Ser Ser Asp Tyr Vai Lys

585590

Ala Met Phe Leu Gly Trp Vai Glu 605

Glu Thr Ser Glu Vai Ser Thr Thr 620

He He Pro Tyr He Gly Pro Ala

635640

Lys Asp Asp Phe Vai Gly Ala Leu

650655

Leu Glu Phe He Pro Glu He Ala

665670

Leu Vai Ser Tyr He Ala Asn Lys 685

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PCT/GB97/02273

865 870 (2) INFORMATION FOR SEQ ID NO: 9:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2628 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE:

(A) NAME/KEY: CDS (B) LOCATION:!..2628 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:

WO 98/07864

PCT/GB97/02273

WO 98/07864

PCT/GB97/02273

AAA TTT GAT AAG TTA Lys Phe Asp Lys Leu 340

AAT TTT GTT AAG TTT Asn Phe Vai Lys Phe 355

TTT GAT AAA GCC GTA

Phe Asp Lys Ala Vai 370

ACA ATA TAT GAT GGA Thr He Tyr Asp Gly 385

TTT AAT GGT CAA AAT Phe Asn Gly Gin Asn 405

AAA AAT TTT ACT GGA Lys Asn Phe Thr Gly 420

GGG ATA ATA ACT TCT Gly lie lie Thr Ser 435

AGC GCT GAT GGG GCA

Ser Ala Asp Gly Ala 450

GAC TTG TTT TTT AGT Asp Leu Phe Phe Ser 465

AAA GGA GAA GAA ATT Lys Gly Glu Glu lie 485

AAT ATT AGT TTA GAT Asn lie Ser Leu Asp 500

GAT AAT GAA CCT GAA

Asp Asn Glu Pro Glu 515

ATA GGC CAA TTA GAA lie Gly Gin Leu Glu .530

AAA AAG TAT GAG TTA Lys Lys Tyr Glu Leu 545

CAA GAA TTT GAA CAT Gin Glu Phe Glu His 565

AAC GAA GCA TTA TTA Asn Glu Ala Leu Leu 580

GAC TAT GTA AAG AAA

Asp Tyr Vai Lys Lys 595

TTG TTT GAA TTT Leu Phe Glu Phe 425

AAA ACT AAA TCA Lys Thr Lys Ser 440

TTA AAT GAT TTA Leu Asn Asp Leu 455

CCT TCA GAA GAT Pro Ser Glu Asp 470

ACA TCT GAT ACT Thr Ser Asp Thr

TTA ATA CAA CAA Leu lie Gin Gin 505

AAT ATT TCA ATA Asn lie Ser lie 520

CTT ATG CCT AAT Leu Met Pro Asn 535

GAT AAA TAT ACT Asp Lys Tyr Thr 550

GGT AAA TCT AGG Gly Lys Ser Arg

AAT CCT AGT CGT Asn Pro Ser Arg 585

GTT AAT AAA GCT Vai Asn Lys Ala 600

TAC AAA ATG TTA Tyr Lys Met Leu 345

TTT AAA GTA CTT Phe Lys Vai Leu 360

TTT AAG ATA AAT Phe Lys He Asn 375

TTT AAT TTA AGA Phe Asn Leu Arg 390

ACA GAA ATT AAT Thr Glu lie Asn

- 56 ACA GAG ATT TAC ACA Thr Glu He Tyr Thr 350

AAC AGA AAA ACA TAT Asn Arg Lys Thr Tyr 365

ATA GTA CCT AAG GTA He Vai Pro Lys Vai 380

AAT ACA AAT TTA GCA

Asn Thr Asn Leu Ala 395

AAT ATG AAT TTT ACT Asn Met Asn Phe Thr 410

TAT AAG TTG CTA TGT Tyr Lys Leu Leu Cys

430

TTA GAT AAA GGA TAC

Leu Asp Lys Gly Tyr 445

TGT ATC AAA GTT AAT

Cys lie Lys Vai Asn 460

AAT TTT ACT AAT GAT

Asn Phe Thr Asn Asp 475

AAT ATA GAA GCA GCA

Asn lie Glu Ala Ala 490

TAT TAT TTA ACC TTT

Tyr Tyr Leu Thr Phe 510

GAA AAT CTT TCA AGT

Glu Asn Leu Ser Ser 525

ATA GAA AGA TTT CCT

He Glu Arg Phe Pro 540

ATG TTC CAT TAT CTT

Met Phe His Tyr Leu 555

ATT GCT TTA ACA AAT He Ala Leu Thr Asn 570

GTT TAT ACA TTT TTT

Vai Tyr Thr Phe Phe 590

ACG GAG GCA GCT ATG Thr Glu Ala Ala Met 605

GAG GAT 1056

Glu Asp

TTG AAT 1104

Leu Asn

AAT TAC 1152

Asn Tyr

GCA AAC 1200

Ala Asn

400

AAA CTA 1248

Lys Leu

415 '

GTA AGA 1296

Vai Arg

AAT AAG 1344

Asn Lys

AAT TGG 1392

Asn Trp

CTA AAT 1440 Leu Asn 480

GAA GAA 1488

Glu Glu 495

AAT TTT 1536

Asn Phe

GAC ATT 1584

Asp lie

AAT GGA 1632

Asn Gly

CGT GCT 1680

Arg Ala

560

TCT GTT 1728

Ser Vai

575

TCT TCA 1776

Ser Ser

TTT TTA 1824 Phe Leu

WO 98/07864

PCT/GB97/02273

865 870 875

WO 98/07864

PCT/GB97/02273

- 58 (2) INFORMATION FOR SEQ ID NO: 10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 876 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10:

100 105 no

WO 98/07864

PCT/GB97/02273

He Gly Pro Ala Leu Asn He Gly Asn Met Leu Tyr Lys Asp Asp Phe

645 650 655

WO 98/07864

PCT/GB97/02273

<td> (2)</td><td colspan="3"> INFORMATION</td><td> FOR</td><td> SEQ</td><td colspan="3"> ID NO: 11:</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2637 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE.TYPE: DNA (genomic) (ix) FEATURE:

(A) NAME/KEY : CDS (B) LOCATION:!. .2637 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:

ATG CAG TTC GTG AAC AAG CAG TTC AAC TAT AAG GAC CCT GTA AAC GGT

Met Gin Phe Vai Asn Lys Gin Phe Asn Tyr Lys Asp Pro Vai Asn Gly 1 5 1015

GTT GAC ATT GCC TAC ATC AAA ATT CCA AAC GCC GGC CAG ATG CAGCCG

Vai Asp He Ala Tyr He Lys He Pro Asn Ala Gly Gin Met GinPro

2530

WO 98/07864

GTG AAG GCT TTC AAG ATT CAT AAC Vai Lys Ala Phe Lys He His Asn 3540

GAT ACA TTT ACG AAC CCG GAA GAA Asp Thr Phe Thr Asn Pro Glu Glu 5055

GCA AAG CAG GTG CCA GTT TCA TAC

Ala Lys Gin Vai Pro Vai Ser Tyr 6570

GAC AAC GAG AAG GAT AAC TAC CTG

Asp Asn Glu Lys Asp Asn Tyr Leu 85

CGT ATT TAT TCC ACT GAC CTG GGC

Arg lie Tyr Ser Thr Asp Leu Gly 100

CGC GGA ATC CCA TTT TGG GGT GGC

Arg Gly lie Pro Phe Trp Gly Gly 115120

GTT ATT GAC ACT AAC TGC ATT AAC Vai lie Asp Thr Asn Cys lie Asn 130135

AGA TCT GAA GAA CTT AAC CTC GTA Arg Ser Glu Glu Leu Asn Leu Vai 145150

ATC CAG TTT GAG TGC AAG AGC TTT lie Gin Phe Glu Cys Lys Ser Phe

165

CGT AAC GGT TAC GGC TCT ACT CAG

Arg Asn Gly Tyr Gly Ser Thr Gin 180

ACG TTC GGT TTC GAG GAG AGC CTG

Thr Phe Gly Phe Glu Glu Ser Leu 195200

GGT GCA GGC AAG TTC GCA ACT GAT

Gly Ala Gly Lys Phe Ala Thr Asp 210215

CTG ATC CAC GCC GGT CAT CGT CTG

Leu lie His Ala Gly His Arg Leu 225230

CGC GTG TTC AAG GTT AAC ACC AAC

Arg Vai Phe Lys Vai Asn Thr Asn 245

GAA GTA AGC TTC GAG GAA CTG CGC

Glu Vai Ser Phe Glu Glu Leu Arg 260

TTT ATC GAC AGC TTG CAG GAG AAC

Phe lie Asp Ser Leu Gin Glu Asn 275 280

AAG TTT AAA GAT ATT GCA AGT ACA Lys Phe Lys Asp lie Ala Ser Thr

290 295

PCT/GB97/02273

- 61 AAA ATC TGG GTT ATT CCG GAA CGC144

Lys He Trp Vai lie Pro Glu Arg

GGA GAC TTG AAC CCG CCG CCG GAA192

Gly Asp Leu Asn Pro Pro Pro Glu

TAC GAT TCA ACC TAT CTG AGC ACA240

Tyr Asp Ser Thr Tyr Leu Ser Thr 7580

AAG GGA GTG ACC AAA TTA TTC GAG2 88

Lys Gly Vai Thr Lys Leu Phe Glu 9095

CGT ATG CTG CTG ACC TCA ATC GTC

Arg Met Leu Leu Thr Ser lie Vai 105110

AGT ACC ATT GAC ACG GAG TTG AAG384

Ser Thr lie Asp Thr Glu Leu Lys

125

GTG ATC CAA CCA GAC GGT AGC TAC4 32

Vai lie Gin Pro Asp Gly Ser Tyr

140

ATC ATC GGG CCC TCC GCG GAC ATT4 80

He He Gly Pro Ser Ala Asp lie

155

GGC CAC GAA GTG TTG AAC CTG ACG528

Gly His Glu Vai Leu Asn Leu Thr

170175

TAC ATT CGT TTC AGC CCA GAC TTC576

Tyr lie Arg Phe Ser Pro Asp Phe

18519Q

GAG GTT GAT ACC AAC CCG CTG TTG624

Glu Vai Asp Thr Asn Pro Leu Leu

205

CCA GCG GTG ACC CTG GCA CAC GAG672

Pro Ala Vai Thr Leu Ala His Glu

220

TAT GGC ATT GCG ATT AAC CCG AAC720

Tyr Gly lie Ala lie Asn Pro Asn

235240

GCC TAC TAC GAG ATG AGT GGT TTA768

Ala Tyr Tyr Glu Met Ser Gly Leu

250255

ACG TTC GGT GGC CAT GAT GCG AAG816

Thr Phe Gly Gly His Asp Ala Lys

265270

GAG TTC CGT CTG TAC TAC TAC AAC864

Glu Phe Arg Leu Tyr Tyr Tyr Asn

285

CTG AAC AAG GCT AAG TCC ATT GTG912

Leu Asn Lys Ala Lys Ser He Vai

300

WO 98/07864

PCT/GB97/02273

565 570 575

WO 98/07864

PCT/GB97/02273

- 63 AAT TCT GTT AAC GAA GCA TTA TTA AAT CCT AGT CGT GTT TAT ACA TTT Asn Ser Vai Asn Glu Ala Leu Leu Asn Pro Ser Arg Vai Tyr Thr Phe 580 585590

TTT TCT TCA GAC TAT GTA AAG AAA GTT AAT AAA GCT ACG GAG GCAGCT

Phe Ser Ser Asp Tyr Vai Lys Lys Vai Asn Lys Ala Thr Glu AlaAla 595 600

ATG TTT TTA GGC TGG GTA GAA CAA TTA GTA TAT GAT TTT ACC GATGAA

Met Phe Leu Gly Trp Vai Glu Gin Leu Vai Tyr Asp Phe Thr AspGlu 610 615620

1776

1824

1872

ACT Thr 625

AGC

Ser

GAA

Glu

GTA

Vai

AGT

Ser

ACT Thr 630

ACG

Thr

GAT

Asp

AAA

Lys

ATT lie

GCG Ala 635

GAT

Asp

ATA

He

ACT

Thr

ATA lie

ATT lie 640

1920

ATT

He

CCA

Pro

TAT

Tyr

ATA

He

GGA

Gly 645

CCT

Pro

GCT

Ala

TTA

Leu

AAT

Asn

ATA He 650

GGT

Gly

AAT

Asn

ATG

Met

TTA

Leu

TAT

Tyr 655

AAA

Lys

1968

GAT

Asp

GAT

Asp

TTT

Phe

GTA

Vai 660

GGT

Gly

GCT

Ala

TTA

Leu

ATA He

TTT

Phe

665

TCA

Ser

GGA

Gly

GCT

Ala

GTT

Vai

ATT

He 670

CTG

Leu

TTA

Leu

2016

GAA TTT ATA CCA GAG ATT GCA ATA CCT GTA TTA GGT ACT TTT GCA CTT

Glu Phe He Pro Glu He Ala He Pro Vai Leu Gly Thr Phe Ala Leu

675 680 685

GTA TCA TAT ATT GCG AAT AAG GTT CTA ACC GTT CAA ACA ATA GAT AAT

Vai Ser Tyr He Ala Asn Lys Vai Leu Thr Vai Gin Thr He Asp Asn

690 695 700

2064

2112

GCT TTA AGT AAA AGA AAT GAA AAA TGG GAT GAG GTC TAT AAA TAT ATA Ala Leu Ser Lys Arg Asn Glu Lys Trp Asp Glu Vai Tyr Lys Tyr He 705 71θ 715 720

2160

2208

2256

2304

2352

2400

2448

2496

2544

WO 98/07864

PCT/GB97/02273

<td> Asp Asn Gin Arg Leu Leu Ser Thr Phe Thr 865 870 (2) INFORMATION FOR SEQ ID NO: 12:</td><td> Glu Tyr lie Lys * 875</td>

(i) SEQUENCE

GTA

Vai

2592

2637

CHARACTERISTICS :

(A) LENGTH: 879 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12

Arg

Vai

Phe

Lys

Vai 245

Asn

Thr

Asn

Ala

Tyr

250

Tyr

Glu

Met

Ser

Gly 255

Leu

WO 98/07864

PCT/GB97/02273

Glu Vai Ser Phe Glu Glu Leu Arg 260

Phe He Asp Ser Leu Gin Glu Asn

275280

Lys Phe Lys Asp lie Ala Ser Thr 290295

Gly Thr Thr Ala Ser Leu Gin Tyr

305310

Tyr Leu Leu Ser Glu Asp Thr Ser 325

Lys Phe Asp Lys Leu Tyr Lys Met 340

Asn Phe Vai Lys Phe Phe Lys Vai

355360

Phe Asp Lys Ala Vai Phe Lys lie 370375

Thr lie Tyr Asp Gly Phe Asn Leu

385390

Phe Asn Gly Gin Asn Thr Glu He 405

Lys Asn Phe Thr Gly Leu Phe Glu 420

Gly lie lie Thr Ser Lys Thr Lys

435440 lie Glu Gly Arg Cys Asp Gly Ala

450455

Asn Asn Trp Asp Leu Phe Phe Ser

465470

Asp Leu Asn Lys Gly Glu Glu He 485

Ala Glu Glu Asn lie Ser Leu Asp 500

Phe Asn Phe Asp Asn Glu Pro Glu

515520

Ser Asp lie lie Gly Gin Leu Glu

530535

Pro Asn Gly Lys Lys Tyr Glu Leu

545550

Leu Arg Ala Gin Glu Phe Glu His

565

Asn Ser Vai Asn Glu Ala Leu Leu 580

Phe Ser Ser Asp Tyr Vai Lys Lys

595 600

- 65 Thr Phe Gly Gly His Asp Ala Lys 265 270

Glu Phe Arg Leu Tyr Tyr Tyr Asn 285

Leu Asn Lys Ala Lys Ser He Vai 300

Met Lys Asn Vai Phe Lys Glu Lys

315320

Gly Lys Phe Ser Vai Asp Lys Leu 330335

Leu Thr Glu lie Tyr Thr Glu Asp

345350

Leu Asn Arg Lys Thr Tyr Leu Asn 365

Asn lie Vai Pro Lys Vai Asn Tyr 380

Arg Asn Thr Asn Leu Ala Ala Asn

395400

Asn Asn Met Asn Phe Thr Lys Leu 410415

Phe Tyr Lys Leu Leu Cys Vai Arg

425430

Ser Leu Asp Lys Gly Tyr Asn Lys 445

Leu Asn Asp Leu Cys lie Lys Vai 460

Pro Ser Glu Asp Asn Phe Thr Asn

475480

Thr Ser Asp Thr Asn lie Glu Ala

490495

Leu He Gin Gin Tyr Tyr Leu Thr 5055io

Asn lie Ser He Glu Asn Leu Ser

525

Leu Met Pro Asn He Glu Arg Phe 540

Asp Lys Tyr Thr Met Phe His Tyr 555560

Gly Lys Ser Arg He Ala Leu Thr 570575

Asn Pro Ser Arg Vai Tyr Thr Phe 585590

Vai Asn Lys Ala Thr Glu Ala Ala 605

WO 98/07864

PCT/GB97/02273

<td> (2)</td><td colspan="3"> INFORMATION</td><td> FOR</td><td> SEQ</td><td colspan="3"> ID NO: 13:</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2862 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE :

(A) NAME/KEY: CDS (B) LOCATION:!..2862 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13:

WO 98/07864

PCT/GB97/02273

WO 98/07864

PCT/GB97/02273

- 68 TTT ATC GAC AGC TTG CAG GAG AAC GAG TTC CGT CTG TAC TAC TAC AAC

Phe He Asp Ser Leu Gin Glu Asn Glu Phe Arg Leu Tyr Tyr Tvr Asn

275 280285

AAG TTT AAA GAT ATT GCA AGT ACA CTG AAC AAG GCT AAG TCC ATTGTG

Lys Phe Lys Asp lie Ala Ser Thr Leu Asn Lys Ala Lys Ser HeVai

290 295300

GGT ACC ACT GCT TCA TTA CAG TAT ATG AAA AAT GTT TTT AAA GAGAAA

Gly Thr Thr Ala Ser Leu Gin Tyr Met Lys Asn Vai Phe Lys GluLvs 305 310 315320

TAT CTC CTA TCT GAA GAT ACA TCT GGA AAA TTT TCG GTA GAT AAATTA

Tyr Leu Leu Ser Glu Asp Thr Ser Gly Lys Phe Ser Vai Asp LysLeu

325 330335

AAA TTT GAT AAG TTA TAC AAA ATG TTA ACA GAG ATT TAC ACA GAGGAT

Lys Phe Asp Lys Leu Tyr Lys Met Leu Thr Glu He Tyr Thr GluAsn 340 345350

AAT TTT GTT AAG TTT TTT AAA GTA CTT AAC AGA AAA ACA TAT TTGAAT

Asn Phe Vai Lys Phe Phe Lys Vai Leu Asn Arg Lys Thr Tyr LeuAsn

355 360355

TTT GAT AAA GCC GTA TTT AAG ATA AAT ATA GTA CCT AAG GTA AATTAC

Phe Asp Lys Ala Vai Phe Lys He Asn He Vai Pro Lys Vai AsnTvr

370 375380

ACA ATA TAT GAT GGA TTT AAT TTA AGA AAT ACA AAT TTA GCA GCAAAC

Thr He Tyr Asp Gly Phe Asn Leu Arg Asn Thr Asn Leu Ala AlaAsn

385 390 395

TTT AAT GGT CAA AAT ACA GAA ATT AAT AAT ATG AAT TTT ACT AAACTA

Phe Asn Gly Gin Asn Thr Glu lie Asn Asn Met Asn Phe Thr LysLeu

405 410415

AAA AAT TTT ACT GGA TTG TTT GAA TTT TAT AAG TTG CTA TGT GTAAGA

Lys Asn Phe Thr Gly Leu Phe Glu Phe Tyr Lys Leu Leu Cys VaiArg

420 425430

GGG ATA ATA ACT TCT AAA ACT AAA TCA TTA GAT AAA GGA TAC AAT AAG

Gly He He Thr Ser Lys Thr Lys Ser Leu Asp Lys Gly Tyr Asn Lys

435 440445

ATC GAA GGT CGT TGC GAT GGG GCA TTA AAT GAT TTA TGT ATC AAA GTT

He Glu Gly Arg Cys Asp Gly Ala Leu Asn Asp Leu Cys He Lys Vai

450 455460

AAT AAT TGG GAC TTG TTT TTT AGT CCT TCA GAA GAT AAT TTT ACT AAT

Asn Asn Trp Asp Leu Phe Phe Ser Pro Ser Glu Asp Asn Phe Thr Asn

465 470 475480

GAT CTA AAT AAA GGA GAA GAA ATT ACA TCT GAT ACT AAT ATA GAA GCA

Asp Leu Asn Lys Gly Glu Glu He Thr Ser Asp Thr Asn He Glu Ala

485 490495

GCA GAA GAA AAT ATT AGT TTA GAT TTA ATA CAA CAA TAT TAT TTA ACC

Ala Glu Glu Asn He Ser Leu Asp Leu lie Gin Gin Tyr Tyr Leu Thr

500 505510

TTT AAT TTT GAT AAT GAA CCT GAA AAT ATT TCA ATA GAA AAT CTTTCA

Phe Asn Phe Asp Asn Glu Pro Glu Asn lie Ser He Glu Asn LeuSer

515 520525

AGT GAC ATT ATA GGC CAA TTA GAA CTT ATG CCT AAT ATA GAA AGATTT

Ser Asp lie lie Gly Gin Leu Glu Leu Met Pro Asn He Glu ArgPhe

530 535540

864

912

960

1008

1056

1104

1152

1200

1248

1296

1344

1392

1440

1488

1536

1584

1632

WO 98/07864

PCT/GB97/O2273

805 810 815

WO 98/07864

PCT/GB97/02273

<td colspan="5"> Cys Phe Arg Ser Cys Asp Leu Arg Arg Leu Glu 930 935 CTC AAG CCT GCC AAG TCA GCT GAA GCT TAG Leu Lys Pro Ala Lys Ser Ala Glu Ala * 945 950 (2) INFORMATION FOR SEQ ID NO: 14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 954 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14</td><td> Met Tyr 940</td><td> Cys Ala Pro</td><td> 2862</td>

Arg He Tyr Ser Thr Asp Leu Gly Arg Met Leu Leu Thr Ser lie Vai

100 105 110

WO 98/07864

PCT/GB97/02273

He Glu Gly Arg Cys Asp Gly Ala Leu Asn Asp Leu Cys He Lys Val 450 455 460

WO 98/07864

PCT/GB97/02273

- 72 Asn Asn Trp Asp Leu Phe Phe Ser Pro Ser Glu Asp Asn Phe Thr Asn 455 470 475 480

Asp Leu Asn Lys Gly Glu Glu He Thr Ser Asp Thr Asn He Glu Ala

485 490 4Ως

Ala Glu Glu Asn He Ser Leu Asp 500

Phe Asn Phe Asp Asn Glu Pro Glu

515520

Ser Asp lie He Gly Gin Leu Glu

530535

Pro Asn Gly Lys Lys Tyr Glu Leu

545550

Leu Arg Ala Gin Glu Phe Glu His

565

Asn Ser Vai Asn Glu Ala Leu Leu 580

Phe Ser Ser Asp Tyr Vai Lys Lys

595600

Met Phe Leu Gly Trp Vai Glu Gin

610615

Thr Ser Glu Vai Ser Thr Thr Asp

625630

He Pro Tyr He Gly Pro Ala Leu

645

Asp Asp Phe Vai Gly Ala Leu He 660

Glu Phe lie Pro Glu He Ala He

675680

Vai Ser Tyr lie Ala Asn Lys Vai

690695

Ala Leu Ser Lys Arg Asn Glu Lys

705710

Vai Thr Asn Trp Leu Ala Lys Vai 725

Lys Lys Met Lys Glu Ala Leu Glu 740 lie lie Asn Tyr Gin Tyr Asn Gin

755760 lie Asn Phe Asn lie Asp Asp Leu

770775

Asn Lys Ala Met lie Asn lie Asn

785790

Ser Tyr Leu Met Asn Ser Met lie 805

Leu lie Gin Gin Tyr Tyr Leu Thr 505 510

Asn lie Ser He Glu Asn Leu Ser 525

Leu Met Pro Asn He Glu Arg Phe 540

Asp Lys Tyr Thr Met Phe His Tyr

555560

Gly Lys Ser Arg He Ala Leu Thr 570575

Asn Pro Ser Arg Vai Tyr Thr Phe 58559o

Vai Asn Lys Ala Thr Glu Ala Ala '605

Leu Vai Tyr Asp Phe Thr Asp Glu 620

Lys He Ala Asp He Thr He He

635640

Asn lie Gly Asn Met Leu Tyr Lys

650655

Phe Ser Gly Ala Vai He Leu Leu

665670

Pro Vai Leu Gly Thr Phe Ala Leu 685

Leu Thr Vai Gin Thr He Asp Asn 700

Trp Asp Glu Vai Tyr Lys Tyr He

715720

Asn Thr Gin He Asp Leu He Arg 730735

Asn Gin Ala Glu Ala Thr Lys Ala

745750

Tyr Thr Glu Glu Glu Lys Asn Asn 765

Ser Ser Lys Leu Asn Glu Ser He 780

Lys Phe Leu Asn Gin Cys Ser Vai

795 800

Pro Tyr Gly Vai Lys Arg Leu Glu

810 815

WO 98/07864

PCT/GB97/02273

- 73 Asp Phe Asp Ala Ser Leu Lys Asp Ala Leu Leu Lys Tyr He Tyr Asn 820 825 830yμ

Asn Arg Gly Thr Leu He Gly Gin Vai Asp Arg Leu Lys Asp LysVai

885 840 845

Asn Asn Thr Leu Ser Thr Asp He Pro Phe Gin Leu Ser Lys Tyr Vai 850 855860

Asp Asn Gin Arg Leu Leu Ser Thr Phe Thr Glu Tyr He Lys SerAra 865 870 875

Pro Gly Pro Glu Thr Leu Cys Gly Ala Glu Leu Vai Asp Ala LeuGin

885 890895

Phe Vai Cys Gly Asp Arg Gly phe Tyr Phe Asn Lys Pro Thr Gly Tyr 900 905gio

Gly Ser Ser Ser Arg Arg Ala Pro Gin Thr Gly He Vai Asp Glu Cys 915 920925

Cys Phe Arg Ser Cys Asp Leu Arg Arg Leu Glu Met Tyr Cys Ala Pro 930 935 94o

Leu Lys Pro Ala Lys Ser Ala Glu Ala * 945950 (2) INFORMATION FOR SEQ ID NO: 15:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2724 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE:· (A) NAME/KEY: CDS (B) LOCATION:!..2724 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15:

ATG CAG TTC GTG AAC AAG CAG TTC AAC TAT AAG GAC CCT GTA AAC GGT Met Gin Phe Vai Asn Lys Gin Phe Asn Tyr Lys Asp Pro Vai Asn Gly 15 10is

GTT GAC ATT GCC TAC ATC AAA ATT CCA AAC GCC GGC CAG ATG CAGCCG

Vai Asp He Ala Tyr He Lys He Pro Asn Ala Gly Gin Met GinPro 20 2530

GTG AAG GCT TTC AAG ATT CAT AAC AAA ATC TGG GTT ATT CCG GAACGC

Vai Lys Ala Phe Lys lie His Asn Lys He Trp Vai lie Pro GluArq

4045

GAT ACA TTT ACG AAC CCG GAA GAA GGA GAC TTG AAC CCG CCG CCGGAA

Asp Thr Phe Thr Asn Pro Glu Glu Gly Asp Leu Asn Pro Pro ProGlu

5560

GCA AAG CAG GTG CCA GTT TCA TAC TAC GAT TCA ACC TAT CTG AGCACA

Ala Lys Gin Vai Pro Vai Ser Tyr Tyr Asp Ser Thr Tyr Leu SerThr 65 70 7580

GAC AAC GAG AAG GAT AAC TAC CTG AAG GGA GTG ACC AAA TTA TTCGAG

Asp Asn Glu Lys Asp Asn Tyr Leu Lys Gly Vai Thr Lys Leu PheGlu

9095

WO 98/07864

PCT/GB97/02273

CGT ATT TAT TCC ACT GAC CTG GGC CGT

Arg He Tyr Ser Thr Asp Leu Gly Arg

100

CGC GGA ATC CCA TTT TGG GGT GGCAGT

Arg Gly lie Pro Phe Trp Gly GlySer

115120

GTT ATT GAC ACT AAC TGC ATT AACGTG

Vai He Asp Thr Asn Cys He AsnVai

130135

AGA TCT GAA GAA CTT AAC CTC GTAATC

Arg Ser Glu Glu Leu Asn Leu VaiHe

145150

ATC CAG TTT GAG TGC AAG AGC TTT GGC

He Gin Phe Glu Cys Lys Ser Phe Gly

165

CGT AAC GGT TAC GGC TCT ACT CAG TAC

Arg Asn Gly Tyr Gly Ser Thr Gin Tyr

180185

ACG TTC GGT TTC GAG GAG AGC CTGGAG

Thr Phe Gly Phe Glu Glu Ser LeuGlu

195200

GGT GCA GGC AAG TTC GCA ACT GATCCA

Gly Ala Gly Lys Phe Ala Thr AspPro

210215

CTG ATC CAC GCC GGT CAT CGT CTGTAT

Leu lie His Ala Gly His Arg LeuTyr

225230

CGC GTG TTC AAG GTT AAC ACC AACGCC

Arg Vai Phe Lys Vai Asn Thr Asn Ala

245

GAA GTA AGC TTC GAG GAA CTG CGC ACG

Glu Vai Ser Phe Glu Glu Leu Arg Thr

260265

TTT ATC GAC AGC TTG CAG GAG AACGAG

Phe lie Asp Ser Leu Gin Glu AsnGlu

275280

AAG TTT AAA GAT ATT GCA AGT ACACTG

Lys Phe Lys Asp lie Ala Ser ThrLeu

290295

GGT ACC ACT GCT TCA TTA CAG TATATG

Gly Thr Thr Ala Ser Leu Gin TyrMet

305310

TAT CTC CTA TCT GAA GAT ACA TCT GGA

Tyr Leu Leu Ser Glu Asp Thr Ser Gly

325

AAA TTT GAT AAG TTA TAC AAA ATG TTA

Lys Phe Asp Lys Leu Tyr Lys Met Leu

340 345

AAT TTT GTT AAG TTT TTT AAA GTA CTT

Asn Phe Vai Lys Phe Phe Lys Vai Leu

355 360

- 74 ATG CTG CTG ACC TCA ATC GTC

Met Leu Leu Thr Ser He Vai

110

ACC ATT GAC ACG GAG TTG AAG

Thr lie Asp Thr Glu Leu Lys'

125

ATC CAA CCA GAC GGT AGC TAC43;

He Gin Pro Asp Gly Ser Tyr 140

ATC GGG CCC TCC GCG GAC ATT lie Gly Pro Ser Ala Asp He 155160

CAC GAA GTG TTG AAC CTG ACG52 8

His Glu Vai Leu Asn Leu Thr 170175

ATT CGT TTC AGC CCA GAC TTC576 lie Arg Phe Ser Pro Aso Phe

190‘

GTT GAT ACC AAC CCG CTG TTG624

Vai Asp Thr Asn Pro Leu Leu 205

GCG GTG ACC CTG GCA CAC GAG672

Ala Vai Thr Leu Ala His Glu 220

GGC ATT GCG ATT AAC CCG AAC720

Gly He Ala He Asn Pro Asn

235240

TAC TAC GAG ATG AGT GGT TTA768

Tyr Tyr Glu Met Ser Gly Leu 250255

TTC GGT GGC CAT GAT GCG AAG816

Phe Gly Gly His Asp Ala Lys

270

TTC CGT CTG TAC TAC TAC AAC864

Phe Arg Leu Tyr Tyr Tyr Asn .285

AAC AAG GCT AAG TCC ATT GTG912

Asn Lys Ala Lys Ser He Vai 300

AAA AAT GTT TTT AAA GAG AAA960

Lys Asn Vai Phe Lys Glu Lys

315320

AAA TTT TCG GTA GAT AAA TTA 1008

Lys Phe Ser Vai Asp Lys Leu 330335

ACA GAG ATT TAC ACA GAG GAT 1056

Thr Glu lie Tyr Thr Glu Asp

350

AAC AGA AAA ACA TAT TTG AAT1104

Asn Arg Lys Thr Tyr Leu Asn 365

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PCT/GB97/02273

- 75 TTT GAT AAA GCC GTA TTT AAG ATA Phe Asp Lys Ala Vai Phe Lys He 370 375

ACA ATA TAT GAT GGA TTT AAT TTA Thr He Tyr Asp Gly Phe Asn Leu 385 390

TTT AAT GGT CAA AAT ACA GAA ATT Phe Asn Gly Gin Asn Thr Glu He 405

AAA AAT TTT ACT GGA TTG TTT GAA

Lys Asn Phe Thr Gly Leu Phe Glu 420

GGG ATA ATA ACT TCT AAA ACT AAA

Gly He He Thr Ser Lys Thr Lys 435440

ATC GAA GGT CGT TGC GAT GGG GCA lie Glu Gly Arg Cys Asp Gly Ala 450455

AAT AAT TGG GAC TTG TTT TTT AGT Asn Asn Trp Asp Leu Phe Phe Ser 465470

GAT CTA AAT AAA GGA GAA GAA ATT

Asp Leu Asn Lys Gly Glu Glu He 485

GCA GAA GAA AAT ATT AGT TTA GAT

Ala Glu Glu Asn lie Ser Leu Asp 500

TTT AAT TTT GAT AAT GAA CCT GAA

Phe Asn Phe Asp Asn Glu Pro Glu 515520

AGT GAC ATT ATA GGC CAA TTA GAA

Ser Asp He He Gly Gin Leu Glu 530535

CCT AAT GGA AAA AAG TAT GAG TTA Pro Asn Gly Lys Lys Tyr Glu Leu 545550

CTT CGT GCT CAA GAA TTT GAA CAT

Leu Arg Ala Gin Glu Phe Glu His 565

AAT TCT GTT AAC GAA GCA TTA TTA

Asn Ser Vai Asn Glu Ala Leu Leu 580

TTT TCT TCA GAC TAT GTA AAG AAA

Phe Ser Ser Asp Tyr Vai Lys Lys 595600

ATG TTT TTA GGC TGG GTA GAA CAA

Met Phe Leu Gly Trp Vai Glu Gin 610615

ACT AGC GAA GTA AGT ACT ACG GAT Thr Ser Glu Vai Ser Thr Thr Asp 625630

AAT ATA GTA CCT AAG GTA AAT TAC Asn lie Vai Pro Lys Vai Asn Tyr 380

AGA AAT ACA AAT TTA GCA GCA AAC

Arg Asn Thr Asn Leu Ala Ala Asn 395400

AAT AAT ATG AAT TTT ACT AAA CTA Asn Asn Met Asn Phe Thr Lys Leu 410415

TTT TAT AAG TTG CTA TGT GTA AGA Phe Tyr Lys Leu Leu Cys Vai Arq 425430

TCA TTA GAT AAA GGA TAC AAT AAG

Ser Leu Asp Lys Gly Tyr Asn Lys 445

TTA AAT GAT TTA TGT ATC AAA GTT

Leu Asn Asp Leu Cys He Lys Vai 460

CCT TCA GAA GAT AAT TTT ACT AAT

Pro Ser Glu Asp Asn Phe Thr Asn 475480

ACA TCT GAT ACT AAT ATA GAA GCA Thr Ser Asp Thr Asn He Glu Ala 490495

TTA ATA CAA CAA TAT TAT TTA ACC Leu He Gin Gin Tyr Tyr Leu Thr 5055io

AAT ATT TCA ATA GAA AAT CTT TCA

Asn He Ser He Glu Asn Leu Ser 525

CTT ATG CCT AAT ATA GAA AGA TTT

Leu Met Pro Asn He Glu Arg Phe 540

GAT AAA TAT ACT ATG TTC CAT TAT

Asp Lys Tyr Thr Met Phe His Tyr 555560

GGT AAA TCT AGG ATT GCT TTA ACA Gly Lys Ser Arg He Ala Leu Thr

570575

AAT CCT AGT CGT GTT TAT ACA TTT

Asn Pro Ser Arg Vai Tyr Thr Phe 585590

GTT AAT AAA GCT ACG GAG GCA GCT

Vai Asn Lys Ala Thr Glu Ala Ala 605

TTA GTA TAT GAT TTT ACC GAT GAA

Leu Vai Tyr Asp Phe Thr Asp Glu 620

AAA ATT GCG GAT ATA ACT ATA ATT

Lys He Ala Asp He Thr He He 635 640

1152

1200

1248

1296

1344

1392

1440

1488

1536

1584

1632

1680

1728

1776

1824

1872

1920

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PCT/GB97/02273

- 76 CCT GCT TTA AAT ATA GGT AAT ATG TTA TAT AAA Pro Ala Leu Asn He Gly Asn Met Leu Tyr Lys 650655

GCT TTA ATA TTT TCA GGA GCT GTT ATT CTGTTA

Ala Leu He Phe Ser Gly Ala Vai He LeuLeu

66567o

ATT GCA ATA CCT GTA TTA GGT ACT TTT GCACTT

He Ala He Pro Vai Leu Gly Thr Phe AlaLeu

680685

AAT AAG GTT CTA ACC GTT CAA ACA ATA GATAAT

Asn Lys Vai Leu Thr Vai Gin Thr lie AspAsn

6957oo

AAT GAA AAA TGG GAT GAG GTC TAT AAA TATATA

Asn Glu Lys Trp Asp Glu Vai Tyr Lys TyrHe 710 715720

GCA AAG GTT AAT ACA CAG ATT GAT CTA ATAAGA

Ala Lys Vai Asn Thr Gin He Asp Leu HeArg

730735

GCT TTA GAA AAT CAA GCA GAA GCA ACA AAGGCT

Ala Leu Glu Asn Gin Ala Glu Ala Thr LysAla

745750

TAT AAT CAA TAT ACT GAG GAA GAG AAA AAT AAT

Tyr Asn Gin Tyr Thr Glu Glu Glu Lys Asn Asn

1968

2016

2064

2112

2160

2208

2256

2304

ATT CCA TAT ATA GGA

He Pro Tyr He Gly

645

GAT GAT TTT GTA GGT

Asp Asp Phe Vai Gly 660

GAA TTT ATA CCA GAG

Glu Phe lie Pro Glu 675

GTA TCA TAT ATT GCG

Vai Ser Tyr lie Ala 690

GCT TTA AGT AAA AGA

Ala Leu Ser Lys Arg 705

GTA ACA AAT TGG TTA

Vai Thr Asn Trp Leu

725

AAA AAA ATG AAA GAA

Lys Lys Met Lys Glu

740

ATA ATA AAC TAT CAG

He lie Asn Tyr Gin

755

ATT AAT TTT AAT ATT GAT GAT TTA AGT TCG AAA CTT AAT GAG TCT ATA lie Asn Phe Asn He Asp Asp Leu Ser Ser Lys Leu Asn Glu Ser He 770 775780

AAT AAA GCT ATG ATT AAT ATA AAT AAA TTT TTG AAT CAA TGC TCTGTT

Asn Lys Ala Met lie Asn He Asn Lys Phe Leu Asn Gin Cys SerVai 785 790 795800

TCA TAT TTA ATG AAT TCT ATG ATC CCT TAT GGT GTT AAA CGG TTAGAA

Ser Tyr Leu Met Asn Ser Met He Pro Tyr Gly Vai Lys Arg LeuGlu

805 810815

2352

2400

2448

2496

2544

2592

900 905

2640

2688

2724

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PCT/GB97/02273

- 77 (2) INFORMATION FOR SEQ ID NO: 16:

(i) SEQUENCE CHARACTERISTICS :

(A) LENGTH: 908 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION:

Met Gin Phe Vai Asn Lys Gin Phe 15

Vai Asp He Ala Tyr lie Lys He 20

Vai Lys Ala Phe Lys He His Asn

3540

Asp Thr Phe Thr Asn Pro Glu Glu 5055

Ala Lys Gin Vai Pro Vai Ser Tyr 6570

Asp Asn Glu Lys Asp Asn Tyr Leu 85

Arg lie Tyr Ser Thr Asp Leu Gly 100

Arg Gly He Pro Phe Trp Gly Glv

115120

Vai lie Asp Thr Asn Cys He Asn 130135

Arg Ser Glu Glu Leu Asn Leu Vai

145150

He Gin Phe Glu Cys Lys Ser Phe 165

Arg Asn Gly Tyr Gly Ser Thr Gin 180

Thr Phe Gly Phe Glu Glu Ser Leu

195200

Gly Ala Gly Lys Phe Ala Thr Asp

210215

Leu lie His Ala Gly His Arg Leu

225230

Arg Vai Phe Lys Vai Asn Thr Asn 245

Glu Vai Ser Phe Glu Glu Leu Arg 260

Phe He Asp Ser Leu Gin Glu Asn

275 280

Lys Phe Lys Asp lie Ala Ser Thr

290 295

SEQ ID NO: 16:

Asn Tyr Lys Asp Pro Vai Asn Gly 10 15

Pro Asn Ala Gly Gin Met Gin Pro 25 30

Lys He Trp Vai He Pro Glu Arg 45

Gly Asp Leu Asn Pro Pro Pro Glu 60

Tyr Asp Ser Thr Tyr Leu Ser Thr 7880

Lys Gly Vai Thr Lys Leu Phe Glu 9095

Arg Met Leu Leu Thr Ser He Vai

105HO

Ser Thr He Asp Thr Glu Leu Lys 125

Vai He Gin Pro Asp Gly Ser Tyr 140

He He Gly Pro Ser Ala Asp He

155160

Gly His Glu Vai Leu Asn Leu Thr 170175

Tyr He Arg Phe Ser Pro Asp Phe 185190

Glu Vai Asp Thr Asn Pro Leu Leu 205

Pro Ala Vai Thr Leu Ala His Glu 220

Tyr Gly He Ala He Asn Pro Asn

235240

Ala Tyr Tyr Glu Met Ser Gly Leu 250255

Thr Phe Gly Gly His Asp Ala Lys

265270

Glu Phe Arg Leu Tyr Tyr Tyr Asn 285

Leu Asn Lys Ala Lys Ser He Vai

300

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PCT/GB97/02273

He Pro Tyr He Gly Pro Ala Leu Asn He Gly Asn Met Leu Tyr Lys

645 650 655

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PCT/GB97/02273

- 79 Asp Asp Phe Vai Gly Ala Leu He Phe Ser Gly Ala Vai He Leu Leu 660 665g70

Glu Phe lie Pro Glu lie Ala lie Pro Vai Leu Gly Thr Phe Ala Leu 675 680685

Vai Ser Tyr He Ala Asn Lys Vai Leu Thr Vai Gin Thr He Asp Asn 690 695700

Ala Leu Ser Lys Arg Asn Glu Lys Trp Asp Glu Vai Tyr Lys TyrHe 705 710 715720

Vai Thr Asn Trp Leu Ala Lys Vai Asn Thr Gin He Asp Leu HeArg

725 730

Lys Lys Met Lys Glu Ala Leu Glu Asn Gin Ala Glu Ala Thr Lys Ala 740 745750

He He Asn Tyr Gin Tyr Asn Gin Tyr Thr Glu Glu Glu Lys Asn Asn 755 760 765'

He Asn Phe Asn He Asp Asp Leu Ser Ser Lys Leu Asn Glu Ser He 770 775780

Asn Lys Ala Met lie Asn lie Asn Lys Phe Leu Asn Gin Cys SerVai 785 790 795800

Ser Tyr Leu Met Asn Ser Met He Pro Tyr Gly Vai Lys Arg LeuGlu

805 810315

Asp Phe Asp Ala Ser Leu Lys Asp Ala Leu Leu Lys Tyr He Tyr Asp 820 825830

Asn Arg Gly Thr Leu He Gly Gin Vai Asp Arg Leu Lys Asp Lys Vai 835 840845

Asn Asn Thr Leu Ser Thr Asp He Pro Phe Gin Leu Ser Lys Tyr Vai 850 855860

Asp Asn Gin Arg Leu Leu Ser Thr Phe Thr Glu Tyr He Lys SerAra 865 θ70 875880

Pro Gin Ser Lys Vai Lys Arg Gin He Phe Ser Gly Tyr Gin SerAsp

885 890895

He Asp Thr His Asn Arg He Lys Asp Glu Leu * 900905 (2) INFORMATION FOR SEQ ID NO: 17:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3042 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE:

(A) NAME/KEY: CDS (B) LOCATION:!..3042 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17:

ATG CAG TTC GTG AAC AAG CAG TTC AAC TAT AAG GAC CCT GTA AAC GGT

Met Gin Phe Vai Asn Lys Gin Phe Asn Tyr Lys Asp Pro Vai Asn Gly 15 10 15

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PCT/GB97/02273

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PCT/GB97/02273

AAG TTT AAA GAT ATT GCA AGT ACA CTG Lys Phe Lys Asp He Ala Ser Thr Leu 290 295

GGT ACC ACT GCT TCA TTA CAG TAT ATG

Gly Thr Thr Ala Ser Leu Gin Tyr Met

305 310

TAT CTC CTA TCT GAA GAT ACA TCT GGA

Tyr Leu Leu Ser Glu Asp Thr Ser Gly

325

AAA TTT GAT AAG TTA TAC AAA ATG TTA

Lys Phe Asp Lys Leu Tyr Lys Met Leu

340345

AAT TTT GTT AAG TTT TTT AAA GTACTT

Asn Phe Vai Lys Phe Phe Lys VaiLeu

355360

TTT GAT AAA GCC GTA TTT AAG ATAAAT

Phe Asp Lys Ala Vai Phe Lys HeAsn

370375

ACA ATA TAT GAT GGA TTT AAT TTAAGA

Thr lie Tyr Asp Gly Phe Asn LeuArg

385390

TTT AAT GGT CAA AAT ACA GAA ATT AAT

Phe Asn Gly Gin Asn Thr Glu lie Asn

405

AAA AAT TTT ACT GGA TTG TTT GAA TTT

Lys Asn Phe Thr Gly Leu Phe Glu Phe

420425

GGG ATA ATA ACT TCT AAA ACT AAATCA

Gly lie lie Thr Ser Lys Thr LysSer

435440

ATC GAA GGT CGT TGC GAT GGG GCATTA

He Glu Gly Arg Cys Asp Gly AlaLeu

450455

AAT AAT TGG GAC TTG TTT TTT AGTCCT

Asn Asn Trp Asp Leu Phe Phe SerPro

465470

GAT CTA AAT AAA GGA GAA GAA ATT ACA

Asp Leu Asn Lys Gly Glu Glu lie Thr

485

GCA GAA GAA AAT ATT AGT TTA GAT TTA

Ala Glu Glu Asn. lie Ser Leu Asp Leu

500505

TTT AAT TTT GAT AAT GAA CCT GAAAAT

Phe Asn Phe Asp Asn Glu Pro GluAsn

515520

AGT GAC ATT ATA GGC CAA TTA GAACTT

Ser Asp lie lie Gly Gin Leu GluLeu

530535

CCT AAT GGA AAA AAG TAT GAG TTAGAT

Pro Asn Gly Lys Lys Tyr Glu LeuAsp

545550

- 81 AAC AAG GCT AAG TCC ATT GTG912

Asn Lys Ala Lys Ser lie Vai

300

AAA AAT GTT TTT AAA GAG AAA960

Lys Asn Vai Phe Lys Glu Lys

315320

AAA TTT TCG GTA GAT AAA TTA10 08

Lys Phe Ser Vai Asp Lys Leu 330335

ACA GAG ATT TAC ACA GAG GAT 1056 Thr Glu He Tyr Thr Glu Asp

350

AAC AGA AAA ACA TAT TTG AAT1104

Asn Arg Lys Thr Tyr Leu Asn

365

ATA GTA CCT AAG GTA AAT TAC 1152 lie Vai Pro Lvs Vai Asn Tyr

380'

AAT ACA AAT TTA GCA GCA AAC 12 00 Asn Thr Asn Leu Ala Ala Asn

395400

AAT ATG AAT TTT ACT AAA CTA 124 8

Asn Met Asn Phe Thr Lys Leu 410415

TAT AAG TTG CTA TGT GTA AGA 1296

Tyr Lys Leu Leu Cys Vai Arg

430

TTA GAT AAA GGA TAC AAT AAG 1344

Leu Asp Lys Gly Tyr Asn Lys

445

AAT GAT TTA TGT ATC AAA GTT 13 92

Asn Asp Leu Cys He Lys Vai 460

TCA GAA GAT AAT TTT ACT AAT 1440

Ser Glu Asp Asn Phe Thr Asn

475 480

TCT GAT ACT AAT ATA GAA GCA 1488

Ser Asp Thr Asn lie Glu Ala 490 495

ATA CAA CAA TAT TAT TTA ACC 1536 He Gin Gin Tyr Tyr Leu Thr

510

ATT TCA ATA GAA AAT CTT TCA 1584 He Ser He Glu Asn Leu Ser

525

ATG CCT AAT ATA GAA AGA TTT 1632

Met Pro Asn He Glu Arg Phe 540

AAA TAT ACT ATG TTC CAT TAT 1680

Lys Tyr Thr Met Phe His Tyr 555 560

WO 98/07864

PCT/GB97/02273

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PCT/GB97/02273

<td> (2)</td><td colspan="3"> INFORMATION</td><td> FOR</td><td> SEQ</td><td colspan="3"> ID NO: 18:</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td colspan="4"> (i) SEQUENCE</td><td colspan="4"> CHARACTERISTICS :</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td colspan="3"> (A) LENGTH</td><td colspan="3"> :: 1014 amino</td><td colspan="2"> acids</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td colspan="3"> (B) TYPE:</td><td colspan="3"> amino acid</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td colspan="4"> (D) TOPOLOGY:</td><td colspan="2"> linear</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td> (ii)</td><td colspan="4"> MOLECULE TYPE:</td><td colspan="2"> protein</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td> (xi)</td><td colspan="8"> SEQUENCE DESCRIPTION: SEQ ID NO</td><td> i: 18</td><td></td><td></td><td></td><td></td><td></td><td></td>

WO 98/07864

PCT/GB97/02273

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PCT/GB97/02273

500 505 gio

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PCT/GB97/02273

1010 (2) INFORMATION FOR SEQ“ID NO: 19:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3509 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE :

(A) NAME/KEY: CDS (B) LOCATION:!..3509 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19:

WO 98/07864

PCT/GB97/02273

WO 98/07864

PCT/GB97/02273

WO 98/07864

PCT/GB97/02273

WO 98/07864

PCT/GB97/02273

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PCT/GB97/02273

- 91 GGA AAT CCT TTA ATG TAC AAT AAA GAA TAT TAT ATG

Gly Asn Pro Leu Met Tyr Asn Lys Glu Tyr Tyr Met

1090 1095 hoi

TTT AAT GCG GGG Phe Asn Ala Gly

CTA GA

Leu

CCT GTA GGT GAA 3360 Pro Vai Gly Glu 1120

TAT ATA AAT TAT 34 08 Tyr He Asn Tyr

1135

AGA AAG TCA AAT 3456

Arg Lys Ser Asn 1150

GAT TAT ATA TAT 3 504

Asp Tyr He Tyr 1165

3509 (2) INFORMATION FOR SEQ ID NO: 20:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1169 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20:

Met Pro Vai Thr He Asn Asn Phe Asn Tyr Asn Asp 15 io

Pro lie Asp Asn

Phe Gly Pro Gly Pro Vai Leu Asn Glu Asn Glu Thr He Asp lie Gly

165 170 175

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PCT/GB97/02273

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PCT/GB97/02273

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PCT/GB97/02273

Leu (2) INFORMATION FOR SEQ ID NO: 21:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2574 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic)

WO 98/07864

PCT/GB97/02273

- 95 (ix) FEATURE:, (A) NAME/KEY : CDS (B) LOCATION:!..2574 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21:

ATG CCA GTT ACA ATA AAT AAT TTT AAT TAT AAT GAT CCT ATT GAT AAT48

Met Pro Vai Thr He Asn Asn Phe Asn Tyr Asn Asp Pro He Asp Asn 15 1015

AAT AAT ATT ATT ATG ATG GAG CCT CCA TTT GCG AGA GGT ACG GGG AGA96

Asn Asn lie lie Met Met Glu Pro Pro Phe Ala Arg Gly Thr GlyArg 20 2530

TAT TAT AAA GCT TTT AAA ATC ACA GAT CGT ATT TGG ATA ATA CCG GAA144

Tyr Tyr Lys Ala Phe Lys He Thr Asp Arg He Trp He He ProGlu

4045

AGA TAT ACT TTT GGA TAT AAA CCT GAG GAT TTT AAT AAA AGT TCC GGT192

Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser SerGly

55go

ATT TTT AAT AGA GAT GTT TGT GAA TAT TAT GAT CCA GAT TAC TTA AAT240

He Phe Asn Arg Asp Vai Cys Glu Tyr Tyr Asp Pro Asp Tyr LeuAsn

70 75

ACT AAT GAT AAA AAG AAT ATA TTT TTA CAA ACA ATG ATC AAG TTA TTT288

Thr Asn Asp Lys Lys Asn lie Phe Leu Gin Thr Met lie Lys LeuPhe

9095

AAT AGA ATC AAA TCA AAA CCA TTG GGT GAA AAG TTA TTA GAG ATG ATT336

Asn Arg lie Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Metlie

100 105no

ATA AAT GGT ATA CCT TAT CTT GGA GAT AGA CGT GTT CCA CTC GAA GAG384

He Asn Gly He Pro Tyr Leu Gly Asp Arg Arg Vai Pro Leu Glu Glu

115 120125

TTT AAC ACA AAC ATT GCT AGT GTA ACT GTT AAT AAA TTA ATC AGT AAT432

Phe Asn Thr Asn lie Ala Ser Vai Thr Vai Asn Lys Leu He Ser Asn

130 135140

CCA GGA GAA G-TG GAG CGA AAA AAA GGT ATT TTC GCA AAT TTA ATA ATA4 80

Pro Gly Glu Vai Glu Arg Lys Lys Gly He Phe Ala Asn Leu lie He

145 150 155160

TTT GGA CCT GGG CCA GTT TTA AAT GAA AAT GAG ACT ATA GAT ATA GGT528

Phe Gly Pro Gly Pro Vai Leu Asn Glu Asn Glu Thr lie Asp He Gly

165 170175

ATA CAA AAT CAT“TTT GCA TCA AGG GAA GGC TTC GGG GGT ATA ATG CAA576 lie Gin Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly He MetGin

180 185190

ATG AAG TTT TGC CCA GAA TAT GTA AGC GTA TTT AAT AAT GTT CAA GAA624

Met Lys Phe Cys Pro Glu Tyr Vai Ser Vai Phe Asn Asn Vai GinGlu

195 200205

AAC AAA GGC GCA AGT ATA TTT AAT AGA CGT GGA TAT TTT TCA GAT CCA672

Asn Lys Gly Ala Ser lie Phe Asn Arg Arg Gly Tyr Phe Ser AspPro

210 215220

GCC TTG ATA TTA ATG CAT GAA CTT ATA CAT GTT TTA CAT GGA TTA TAT72 0

Ala Leu lie Leu Met His Glu Leu He His Vai Leu His Gly LeuTyr

225 230 235240

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PCT/GB97/02273

- 96 GGC ATT AAA GTA GAT GAT TTA CCA ATT GTA CCA AAT GAA AAA AAA TTT768

Gly He Lys Vai Asp Asp Leu Pro lie Vai Pro Asn Glu Lys Lys Phe

245 250255

TTT ATG CAA TCT ACA GAT GCT ATA CAG GCA GAA GAA CTA TAT ACA TTT816

Phe Met Gin Ser Thr Asp Ala lie Gin Ala Glu Glu Leu Tyr ThrPhe

260 265270

GGA GGA CAA GAT CCC AGC ATC ATA ACT CCT TCT ACG GAT AAA AGT ATC864

Gly Gly Gin Asp Pro Ser He He Thr Pro Ser Thr Asp Lys SerHe

275 280285

TAT GAT AAA GTT TTG CAA AAT TTT AGA GGG ATA GTT GAT AGA CTT AAC912

Tyr Asp Lys Vai Leu Gin Asn Phe Arg Gly He Vai Asp Arg LeuAsn

290 295300

AAG GTT TTA GTT TGC ATA TCA GAT CCT AAC ATT AAT ATT AAT ATA TAT960

Lys Vai Leu Vai Cys He Ser Asp Pro Asn He Asn He Asn HeTyr

305 310 315320

AAA AAT AAA TTT AAA GAT AAA TAT AAA TTC GTT GAA GAT TCT GAG GGA1008

Lys Asn Lys Phe Lys Asp Lys Tyr Lys Phe Vai Glu Asp Ser GluGly

325 330335

AAA TAT AGT ATA GAT GTA GAA AGT TTT GAT AAA TTA TAT AAA AGC TTA1056

Lys Tyr Ser He Asp Vai Glu Ser Phe Asp Lys Leu Tyr Lys SerLeu

340 345350

ATG TTT GGT TTT ACA GAA ACT AAT ATA GCA GAA AAT TAT AAA ATA AAA1104

Met Phe Gly Phe Thr Glu Thr Asn He Ala Glu Asn Tyr Lys HeLys

355 360365

ACT AGA GCT TCT TAT TTT AGT GAT TCC TTA CCA CCA GTA AAA ATA AAA1152

Thr Arg Ala Ser Tyr Phe Ser Asp Ser Leu Pro Pro Vai Lys lieLys

370 375380

AAT TTA TTA GAT AAT GAA ATC TAT ACT ATA GAG GAA GGG TTT AAT ATA1200

Asn Leu Leu Asp Asn Glu He Tyr Thr He Glu Glu Gly Phe AsnHe

385 390 395

TCT GAT AAA GAT ATG GAA AAA GAA TAT AGA GGT CAG AAT AAA GCT ATA124 8

Ser Asp Lys Asp Met Glu Lys Glu Tyr Arg Gly Gin Asn Lys AlaHe

405 410415

AAT AAA CAA GCT TAT GAA GAA ATT AGC AAG GAG CAT TTG GCT GTA TAT1296

Asn Lys Gin Ala Tyr Glu Glu lie Ser Lys Glu His Leu Ala VaiTyr

420 425430

AAG ATA CAA ATG TGT AAA AGT GTT AAA GCT CCA GGA ATA TGT ATT GAT1344

Lys lie Gin Met Cys Lys Ser Vai Lys Ala Pro Gly lie Cys lieAsp

435 440445

GTT GAT AAT GAA GAT TTG TTC TTT ATA GCT GAT AAA AAT AGT TTT TCA1392

Vai Asp Asn Glu Asp Leu Phe Phe lie Ala Asp Lys Asn Ser PheSer

450 455460

GAT GAT TTA TCT AAA AAC GAA AGA ATA GAA TAT AAT ACA CAG AGT AAT144 0

Asp Asp Leu Ser Lys Asn Glu Arg lie Glu Tyr Asn Thr Gin SerAsn

465 470 475480

TAT ATA GAA AAT GAC TTC CCT ATA AAT GAA TTA ATT TTA GAT ACT GAT1488

Tyr He Glu Asn Asp Phe Pro lie Asn Glu Leu He Leu Asp ThrAsp

485 490495

TTA ATA AGT AAA ATA GAA TTA CCA AGT GAA AAT ACA GAA TCA CTT ACT153 6

Leu lie Ser Lys lie Glu Leu Pro Ser Glu Asn Thr Glu Ser LeuThr

500 505510

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PCT/GB97/02273

- 97 GAT TTT AAT GTA GAT GTT CCA GTA TAT GAA AAA CAA CCC GCT ATA AAA1584

Asp Phe Asn Val Asp Val Pro Val Tyr Glu Lys Gin Pro Ala HeLys

515 520525

AAA ATT TTT ACA GAT GAA AAT ACC ATC TTT CAA TAT TTA TAC TCT CAG1632

Lys lie Phe Thr Asp Glu Asn Thr lie Phe Gin Tyr Leu Tyr SerGin

530 535540

ACA TTT CCT CTA GAT ATA AGA GAT ATA AGT TTA ACA TCT TCA TTT GAT1680

Thr Phe Pro Leu Asp lie Arg Asp He Ser Leu Thr Ser Ser PheAsp

545 550 555560

GAT GCA TTA TTA TTT TCT AAC AAA GTT TAT TCA TTT TTT TCT ATG GAT1728

Asp Ala Leu Leu Phe Ser Asn Lys Val Tyr Ser Phe Phe Ser MetAsp

565 570575

TAT ATT AAA ACT GCT AAT AAA GTG GTA GAA GCA GGA TTA TTT GCA GGT1776

Tyr lie Lys Thr Ala Asn Lys Val Val Glu Ala Gly Leu Phe AlaGly

580 585590

TGG GTG AAA CAG ATA GTA AAT GAT TTT GTA ATC GAA GCT AAT AAA AGC1824

Trp Val Lys Gin He Val Asn Asp Phe Val He Glu Ala Asn LysSer

595 600605

AAT ACT ATG GAT AAA ATT GCA GAT ATA TCT CTA ATT GTT CCT TAT ATA1872

Asn Thr Met Asp Lys He Ala Asp lie Ser Leu lie Val Pro TyrHe

610 615620

GGA TTA GCT TTA AAT GTA GGA AAT GAA ACA GCT AAA GGA AAT TTT GAA1920

Gly Leu Ala Leu Asn Val Gly Asn Glu Thr Ala Lys Gly Asn PheGlu

625 630 635640

AAT GCT TTT GAG ATT GCA GGA GCC AGT ATT CTA CTA GAA TTT ATA CCA1968

Asn Ala Phe Glu lie Ala Gly Ala Ser He Leu Leu Glu Phe liePro

645 650655

GAA CTT TTA ATA CCT GTA GTT GGA GCC TTT TTA TTA GAA TCA TAT ATT2016

Glu Leu Leu lie Pro Val Val Gly Ala Phe Leu Leu Glu Ser Tyrlie

660 665670

GAC AAT AAA AAT AAA ATT ATT AAA ACA ATA GAT AAT GCT TTA ACT AAA2064

Asp Asn Lys Asn Lys lie lie Lys Thr lie Asp Asn Ala Leu ThrLys

675 680685

AGA AAT GAA AAA TGG AGT GAT ATG TAC GGA TTA ATA GTA GCG CAA TGG2112

Arg Asn Glu Lys Trp Ser Asp Met Tyr Gly Leu lie Val Ala GinTrp

690 695700

CTC TCA ACA GTT AAT ACT CAA TTT TAT ACA ATA AAA GAG GGA ATG TAT2160

Leu Ser Thr Val Asn Thr Gin Phe Tyr Thr lie Lys Glu Gly MetTyr

705 710 715720

AAG GCT TTA AAT TAT CAA GCA CAA GCA TTG GAA GAA ATA ATA AAA TAC2208

Lys Ala Leu Asn Tyr Gin Ala Gin Ala Leu Glu Glu He He LysTyr

725 730735

AGA TAT AAT ATA TAT TCT GAA AAA GAA AAG TCA AAT ATT AAC ATC GAT2256

Arg Tyr Asn lie Tyr Ser Glu Lys Glu Lys Ser Asn He Asn lieAsp

740 745750

TTT AAT GAT ATA AAT TCT AAA CTT AAT GAG GGT ATT AAC CAA GCT ATA2304

Phe Asn Asp lie Asn Ser Lys Leu Asn Glu Gly He Asn Gin AlaHe

755 760765

GAT AAT ATA AAT AAT TTT ATA AAT GGA TGT TCT GTA TCA TAT TTA ATG2352

Asp Asn lie Asn Asn Phe lie Asn Gly Cys Ser Val Ser Tyr LeuMet

770 775780

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- 98 AAA AAA ATG ATT CCA TTA GCT GTA GAA AAA TTA CTA GAC TTT GAT AAT

Lys Lys Met He Pro Leu Ala Vai Glu Lys Leu Leu Asp Phe Asn Asn

735 790 795800

ACT CTC AAA AAA AAT TTG TTA AAT TAT ATA GAT GAA AAT AAA TTATAT

Thr Leu Lys Lys Asn Leu Leu Asn Tyr He Asp Glu Asn Lys LeuTyr

805 810815

TTG ATT GGA AGT GCA GAA TAT GAA AAA TCA AAA GTA AAT AAA TACTTG

Leu lie Gly Ser Ala Glu Tyr Glu Lys Ser Lys Vai Asn Lys TyrLeu

820 825830

AAA ACC ATT ATG CCG TTT GAT CTT TCA ATA TAT ACC AAT GAT ACAATA

Lys Thr lie Met Pro Phe Asp Leu Ser He Tyr Thr Asn Asp ThrHe

835 840845

CTA ATA GAA ATG TTT AAT AAA TAT AATAGC

Leu lie Glu Met Phe Asn Lys Tyr AsnSer

850855

2400

2448

2496

2544

2574 (2) INFORMATION FOR SEQ ID NO: 22:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 858 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22:

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- 100 -

(2) INFORMATION FOR SEQ ID NO: 23:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1644 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear

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- 101 (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE:

(A) NAME/KEY: CDS (B) LOCATION:1. .1644 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 23:

ATG CCA GTT ACA ATA AAT AAT TTT AAT Met Pro Vai Thr He Asn Asn Phe Asn 15

AAT AAT ATT ATT ATG ATG GAG CCTCCA

Asn Asn lie He Met Met Glu ProPro

2025

TAT TAT AAA GCT TTT AAA ATC ACAGAT

Tyr Tyr Lys Ala Phe Lys He ThrAsp

3540

AGA TAT ACT TTT GGA TAT AAA CCTGAG

Arg Tyr Thr Phe Gly Tyr Lys ProGlu

5055

ATT TTT AAT AGA GAT GTT TGT GAATAT

He Phe Asn Arg Asp Vai Cys GluTyr

6570

ACT AAT GAT AAA AAG AAT ATA TTT TTA

Thr Asn Asp Lys Lys Asn He Phe Leu

AAT AGA ATC AAA TCA AAA CCA TTG GGT

Asn Arg He Lys Ser Lys Pro Leu Gly

100105

ATA AAT GGT ATA CCT TAT CTT GGA'GAT lie Asn Gly lie Pro Tyr Leu Gly Asp

115120

TTT AAC ACA AAC ATT GCT AGT GTAACT

Phe Asn Thr Asn lie Ala Ser VaiThr

130135

CCA GGA GAA GTG GAG CGA AAA AAAGGT

Pro Gly Glu Vai Glu Arg Lys LvsGly

145 150*

TTT GGA CCT GGG CCA GTT TTA AAT GAA

Phe Gly Pro Gly Pro Vai Leu Asn Glu

165

ATA CAA AAT CAT TTT GCA TCA AGG GAA lie Gin Asn His Phe Ala Ser Arg Glu

180185

ATG AAG TTT TGC CCA GAA TAT GTAAGC

Met Lys Phe Cys Pro Glu Tyr VaiSer

195200

AAC AAA GGC GCA AGT ATA TTT AATAGA

Asn Lys Gly Ala Ser lie Phe AsnArg

210215

TAT AAT GAT CCT ATT GAT AAT4 8

Tyr Asn Asp Pro He Asp Asn

1015

TTT GCG AGA GGT ACG GGG AGA96

Phe Ala Arg Gly Thr Gly Arg

CGT ATT TGG ATA ATA CCG GAA144

Arg He Trp lie He Pro Glu

GAT TTT AAT AAA AGT TCC GGT192

Asp Phe Asn Lys Ser Ser Gly

TAT GAT CCA GAT TAC TTA AAT24 0

Tyr Asp Pro Asp Tyr Leu Asn

7580

CAA ACA ATG ATC AAG TTA TTT28 8

Gin Thr Met lie Lys Leu Phe

9095

GAA AAG TTA TTA GAG ATG ATT33 6

Glu Lys Leu Leu Glu Met lie

110

AGA CGT GTT CCA CTC GAA GAG3 84

Arg Arg Vai Pro Leu Glu Glu

125

GTT AAT AAA TTA ATC AGT AAT432

Vai Asn Lys Leu He Ser Asn

140

ATT TTC GCA AAT TTA ATA ATA4 80 lie Phe Ala Asn Leu lie lie

155160

AAT GAG ACT ATA GAT ATA GGT528

Asn Glu Thr lie Asp lie Gly

170175

GGC TTC GGG GGT ATA ATG CAA576

Gly Phe Gly Gly He Met Gin

190

GTA TTT AAT AAT GTT CAA GAA624

Vai Phe Asn Asn Vai Gin Glu

205

CGT GGA TAT TTT TCA GAT CCA672

Arg Gly Tyr Phe Ser Asp Pro

220

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- 102 -

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PCT/GB97/02273

- 103 TTA ATA AGT AAA ATA GAA TTA CCA AGT GAA AAT ACA GAA TCA CTT ACT Leu He Ser Lys He Glu Leu Pro Ser Glu Asn Thr Glu Ser Leu Thr 500 505510

GAT TTT AAT GTA GAT GTT CCA GTA TAT GAA AAA CAA CCC GCT ATAAAA

Asp Phe Asn Vai Asp Vai Pro Vai Tyr Glu Lys Gin Pro Ala HeLys

515 520525

AAA ATT TTT ACA GAT GAA AAT ACC ATC TTT CAA TAT TTA TAC TCTCAG

Lys He Phe Thr Asp Glu Asn Thr He Phe Gin Tyr Leu Tyr SerGin

530 535540

ACA TTT CCT CTA

Thr Phe Pro Leu 545 (2) INFORMATION FOR SEQ ID NO: 24:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 548 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24:·

Met Pro Vai Thr He Asn Asn Phe Asn Tyr Asn Asp Pro He Asp Asn 1 5 1015

Asn Asn lie lie Met Met Glu Pro Pro Phe Ala Arg Gly Thr Gly Ara 20 2530

Tyr Tyr Lys Ala Phe Lys lie Thr Asp Arg He Trp He lie Pro Glu 35 4045

Arg Tyr Thr Phe Gly Tyr Lys Pro Glu Asp Phe Asn Lys Ser Ser Gly 50 5560 lie Phe Asn Arg Asp Vai Cys Glu Tyr Tyr Asp Pro Asp Tyr LeuAsn 65 70 75Θ0

Thr Asn Asp Lys Lys Asn He Phe Leu Gin Thr Met He Lys LeuPhe

9095

Asn Arg He Lys Ser Lys Pro Leu Gly Glu Lys Leu Leu Glu Met He 100 105110

He Asn Gly lie Pro Tyr Leu Gly Asp Arg Arg Vai Pro Leu Glu Glu

H5 120125

Phe Asn Thr Asn He Ala Ser Vai Thr Vai AsrrLys Leu He Ser Asn 130 135140

Pro Gly Glu Vai Glu Arg Lys Lys Gly He Phe Ala Asn Leu HeHe

145 150 155160

Phe Gly Pro Gly Pro Vai Leu Asn Glu Asn Glu Thr He Asp HeGly

165 170175 lie Gin Asn His Phe Ala Ser Arg Glu Gly Phe Gly Gly He Met Gin 180 185190

Met Lys Phe Cys Pro Glu Tyr Vai Ser Vai Phe Asn Asn Vai Gin Glu 195 200205

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- 104 -

Thr Phe Pro Leu 545

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- 105 (2) INFORMATION FOR SEQ ID NO: 25:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2616 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE:

(A) NAME/KEY: CDS (B) LOCATION:!..2616 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25:

ATG CAG TTC GTG AAC AAG CAG TTC

Met Gin Phe Vai Asn Lys Gin Phe

GTT GAC ATT GCC TAC ATC AAAATT

Vai Asp He Ala Tyr He LysHe

GTG AAG GCT TTC AAG ATT CATAAC

Vai Lys Ala Phe Lys lie HisAsn

3540

GAT ACA TTT ACG AAC CCG GAA GAA

Asp Thr Phe Thr Asn Pro Glu Glu 5055

GCA AAG CAG GTG CCA GTT TCA TAC

Ala Lys Gin Vai Pro Vai Ser Tyr 6570

GAC AAC GAG AAG GAT AAC TAC CTG

Asp Asn Glu Lys Asp Asn Tyr Leu 85

CGT ATT TAT TCC ACT GAC CTG GGC

Arg He Tyr Ser Thr Asp Leu Gly 100

CGC GGA ATC CCA TTT TGG GGT GGC

Arg Gly He Pro Phe Trp Gly Gly

115120

GTT ATT GAC ACT AAC TGC ATT AAC

Vai lie Asp Thr Asn Cys lie Asn 130135

AGA TCT GAA GAA CTT AAC CTC GTA

Arg Ser Glu Glu Leu Asn Leu Vai 145150

ATC CAG TTT GAG TGC AAG AGC TTT lie Gin Phe Glu Cys Lys Ser Phe

165

CGT AAC GGT TAC GGC TCT ACT CAG

Arg Asn Gly Tyr Gly Ser Thr Gin 180

AAC TAT AAG GAC CCT GTA AAC GGT4 8

Asn Tyr Lys Asp Pro Vai Asn Gly

1015

CCA AAC GCC GGC CAG ATG CAG CCG96

Pro Asn Ala Gly Gin Met GinPro

2530

AAA ATC TGG GTT ATT CCG GAA CGC144

Lys He Trp Vai lie Pro GluArg

GGA GAC TTG AAC CCG CCG CCG GAA192

Gly Asp Leu Asn Pro Pro ProGlu

TAC GAT TCA ACC TAT CTG AGC ACA240

Tyr Asp Ser Thr Tyr Leu SerThr

7580

AAG GGA GTG ACC AAA TTA TTC GAG28 8

Lys Gly Vai Thr Lys Leu PheGlu

9095

CGT ATG CTG CTG ACC TCA ATC GTC336

Arg Met Leu Leu Thr Ser lieVai

105no

AGT ACC ATT GAC ACG GAG TTG AAG384

Ser Thr He Asp Thr Glu LeuLys

125

GTG ATC CAA CCA GAC GGT AGC TAC432

Vai lie Gin Pro Asp Gly SerTyr

140

ATC ATC GGG CCC TCC GCG GAC ATT480 lie lie Gly Pro Ser Ala Asplie

155160

GGC CAC GAA GTG TTG AAC CTG ACG528

Gly His Glu Vai Leu Asn LeuThr

170175

TAC ATT CGT TTC AGC CCA GAC TTC576

Tyr lie Arg Phe Ser Pro AspPhe

185190

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ACG TTC GGT TTC GAG GAG AGC CTG GAG GTT Thr Phe Gly Phe Glu Glu Ser Leu Glu Vai 195200

GGT GCA GGC AAG TTC GCA ACT GAT CCAGCG

Gly Ala Gly Lys Phe Ala Thr Asp ProAla

210215

CTG ATC CAC GCC GGT CAT CGT CTG TATGGC

Leu He His Ala Gly His Arg Leu TyrGly

225230

CGC GTG TTC AAG GTT AAC ACC AAC GCCTAC

Arg Vai Phe Lys Vai Asn Thr Asn AlaTyr

245250

GAA GTA AGC TTC GAG GAA CTG CGC ACGTTC

Glu Vai Ser Phe Glu Glu Leu Arg ThrPhe

260265

TTT ATC GAC AGC TTG CAG GAG AAC GAGTTC

Phe lie Asp Ser Leu Gin Glu Asn GluPhe

275280

AAG TTT AAA GAT ATT GCA AGT ACA CTGAAC

Lys Phe Lys Asp lie Ala Ser Thr LeuAsn

290295

GGT ACC ACT GCT TCA TTA CAG TAT ATGAAA

Gly Thr Thr Ala Ser Leu Gin Tyr MetLys

305310

TAT CTC CTA TCT GAA GAT ACA TCT GGAAAA

Tyr Leu Leu Ser Glu Asp Thr Ser GlyLys

325330

AAA TTT GAT AAG TTA TAC AAA ATG TTAACA

Lys Phe Asp Lys Leu Tyr Lys Met LeuThr

340345

AAT TTT GTT AAG TTT TTT AAA GTA CTTAAC

Asn Phe Vai Lys Phe Phe Lys Vai LeuAsn

355360

TTT GAT AAA GCC GTA TTT AAG ATA AATATA

Phe Asp Lys Ala Vai Phe Lys lie Asnlie

370375

ACA ATA TAT GAT GGA TTT AAT TTA AGAAAT

Thr He Tyr Asp Gly Phe Asn Leu ArgAsn

385390

TTT AAT GGT CAA AAT ACA GAA ATT AATAAT

Phe Asn Gly Gin Asn Thr Glu He AsnAsn

405410

AAA AAT TTT ACT GGA TTG TTT GAA TTTTAT

Lys Asn Phe Thr Gly Leu Phe Glu PheTyr

420425

GGG ATA ATA ACT TCT AAA ACT AAA TCATTA

Gly lie He Thr Ser Lys Thr Lys SerLeu

435440

GCA TTA AAT GAT TTA TGT ATC AAA GTTAAT

Ala Leu Asn Asp Leu Cys lie Lys VaiAsn

450455

106 GAT ACC AAC CCG CTG TTG

Asp Thr Asn Pro Leu Leu

205

GTG ACC CTG GCA CAC GAG672

Vai Thr Leu Ala His Glu 220

ATT GCG ATT AAC CCG AAC720

He Ala He Asn Pro Asn

235240

TAC GAG ATG AGT GGT TTA768

Tyr Glu Met Ser Gly Leu

255

GGT GGC CAT GAT GCG AAG816

Gly Gly His Asp Ala Lys

270

CGT CTG TAC TAC TAC AAC864

Arg Leu Tyr Tyr Tyr Asn

285

AAG GCT AAG TCC ATT GTG912

Lys Ala Lys Ser He Vai

300

AAT GTT TTT AAA GAG AAA960

Asn Vai Phe Lys Glu Lys

315320

TTT TCG GTA GAT AAA TTA 100 8

Phe Ser Vai Asp Lys Leu

335

GAG ATT TAC ACA GAG GAT 1056

Glu He Tyr Thr Glu Asp

350

AGA AAA ACA TAT TTG AAT 1104

Arg Lys Thr Tyr Leu Asn

365

GTA CCT AAG GTA AAT TAC 1152

Vai Pro Lys Vai Asn Tyr 380

ACA AAT TTA GCA GCA AAC 1200

Thr Asn Leu Ala Ala Asn

395 400

ATG AAT TTT ACT AAA CTA 124 8

Met Asn Phe Thr Lys Leu

415

AAG TTG CTA TGT GTA AGA 1296

Lys Leu Leu Cys Vai Arg

430

GAT AAA GGA TAC AAT AAG 1344

Asp Lys Gly Tyr Asn Lys

445

AAT TGG GAC TTG TTT TTT 13 92

Asn Trp Asp Leu Phe Phe

460

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- 107 -

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- 108 -

<td> He ACA Thr 865 (2)</td><td colspan="7"> Pro Phe Gin Leu Ser Lys Tyr Vai Asp Asn Gin 850 855 860 TTT ACT GAA TAT ATT AAG TAA Phe Thr Glu Tyr He Lys * 870 INFORMATION FOR SEQ ID NO: 26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 872 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26:</td><td> Arg</td><td> Leu Leu</td><td> Ser</td><td> 2616</td>

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Arg Gly He Pro Phe Trp Gly 115

Val lie Asp Thr Asn Cys He

130 135

Arg Ser Glu Glu Leu Asn Leu 145 150 lie Gin Phe Glu Cys Lys Ser

165

Arg Asn Gly Tyr Gly Ser Thr 180

Thr Phe Gly Phe Glu Glu Ser 195

Gly Ala Gly Lys Phe Ala Thr

210 215

Leu lie His Ala Gly His Arg 225 230

Arg Val Phe Lys Val Asn Thr 245

Glu Val Ser Phe Glu Glu Leu

260

Phe lie Asp Ser Leu Gin Glu 275

Lys Phe Lys Asp lie Ala Ser

290 295

Gly Thr Thr Ala Ser Leu Gin 305 310

Tyr Leu Leu Ser Glu Asp Thr

325

Lys Phe Asp Lys Leu Tyr Lys

340

Asn Phe Val Lys Phe Phe Lys 355

Phe Asp Lys Ala Val Phe Lys

370 375

Thr He Tyr Asp Gly Phe Asn

385 390

Phe Asn Gly Gin Asn Thr Glu

405

Lys Asn Phe Thr Gly Leu Phe 420

Gly He lie Thr Ser Lys Thr 435

Ala Leu Asn Asp Leu Cys lie

450 455

- 109 Gly Ser Thr lie Asp Thr Glu Leu Lys 120125

Asn Val He Gin Pro Asp Gly Ser Tyr 140

Val He lie Gly Pro Ser Ala Asp He

155160

Phe Gly His Glu Val Leu Asn Leu Thr 170175

Gin Tyr He Arg Phe Ser Pro Asp Phe 185190

Leu Glu Val Asp Thr Asn Pro Leu Leu

200205

Asp Pro Ala Val Thr Leu Ala His Glu 220

Leu Tyr Gly He Ala He Asn Pro Asn

235240

Asn Ala Tyr Tyr Glu Met Ser Gly Leu 250255

Arg Thr Phe Gly Gly His Asp Ala Lys 265270

Asn Glu Phe Arg Leu Tyr Tyr Tyr Asn

280285

Thr Leu Asn Lys Ala Lys Ser lie Val 300

Tyr Met Lys Asn Val Phe Lys Glu Lys

315320

Ser Gly Lys Phe Ser Val Asp Lys Leu 330335

Met Leu Thr Glu He Tyr Thr Glu Asp

345350

Val Leu Asn Arg Lys Thr Tyr Leu Asn

360365 lie Asn lie Val Pro Lys Val Asn Tyr 380

Leu Arg Asn Thr Asn Leu Ala Ala Asn

395400

He Asn Asn Met Asn Phe Thr Lys Leu 410415

Glu Phe Tyr Lys Leu Leu Cys Val Arg 425430

Lys Ser Leu Asp Lys Gly Tyr Asn Lys

440445

Lys Val Asn Asn Trp Asp Leu Phe Phe 460

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-HO-

805 810 815

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- 111 Asp Ala Leu Leu Lys Tyr He Tyr Asp Asn Arg Gly Thr Leu lie Glv 820 825830

Gin Vai Asp Arg Leu Lys Asp Lys Vai Asn Asn Thr Leu Ser Thr Asd 335 840845

He Pro Phe Gin Leu Ser Lys Tyr Vai Asp Asn Gin Arg Leu Leu Ser 850 855860

Thr Phe Thr Glu Tyr lie Lys *

865870 (2) INFORMATION FOR SEQ ID NO: 27:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2574 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27:·

ATGCCGGTTA CCATCAACAA CTTCAACTAC AACGACCCGA TCGACAACAA CAACATCATC 60 ATGATGGAAC CGCCGTTCGC ACGTGGTACC GGTCGTTACT ACAAGGCTTT CAAGATCACC 120 GACCGTATCT GGATCATCCC GGAACGTTAC ACCTTCGGTT ACAAACCTGA GGACTTCAAC 180 AAGAGTAGCG GGATTTTCAA TCGTGACGTC TGCGAGTACT ATGATCCAGA TTATCTGAAT 240 ACCAACGATA AGAAGAACAT ATTCCTTCAG ACTATGATCA AGTTATTTAA TAGAATCAAA 300 TCAAAACCAT TGGGTGAAAA GTTATTAGAG ATGATTATAA ATGGTATACC TTATCTTGGA 360 GATAGACGTG TTCCACTCGA AGAGTTTAAC ACAAACATTG CTAGTGTAAC TGTTAATAAA 420 TTAATCAGTA ATCCAGGAGA AGTGGAGCGA AAAAAAGGTA TTTTCGCAAA TTTAATAATA 480 TTTGGACCTG GGCCAGTTTT AAATGAAAAT GAGACTATAG ATATAGGTAT ACAAAATCAT 540 TTTGCATCAA GGGAAGGCTT CGGGGGTATA ATGCAAATGA AGTTTTGCCC AGAATATGTA 600 AGCGTATTTA ATAATGTTCA AGAAAACAAA GGCGCAAGTA TATTTAATAG ACGTGGATAT 660 TTTTCAGATC CAGCCTTGAT ATTAATGCAT GAACTTATAC ATGTTTTACA TGGATTATAT 720 GGCATTAAAG TAGATGATTT ACCAATTGTA CCAAATGAAA AAAAATTTTT TATGCAATCT 780 ACAGATGCTA TACAGGCAGA AGAACTATAT ACATTTGGAG GACAAGATCC CAGCATCATA 840 ACTCCTTCTA CGGATAAAAG TATCTATGAT AAAGTTTTGC AAAATTTTAG AGGGATAGTT 900 GATAGACTTA ACAAGGTTTT AGTTTGCATA TCAGATCCTA ACATTAATAT TAATATATAT 960 AAAAATAAAT TTAAAGATAA ATATAAATTC GTTGAAGATT CTGAGGGAAA ATATAGTATA 1020 GATGTAGAAA GTTTTGATAA ATTATATAAA AGCTTAATGT TTGGTTTTAC AGAAACTAAT 1080 ATAGCAGAAA ATTATAAAAT AAAAACTAGA GCTTCTTATT TTAGTGATTC CTTACCACCA 1140 GTAAAAATAA AAAATTTATT AGATAATGAA ATCTATACTA TAGAGGAAGG GTTTAATATA 1200

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- 112 -

<td> TCTGATAAAG</td><td> ATATGGAAAA</td><td> AGAATATAGA</td><td> GGTCAGAATA</td><td> AAGCTATAAA</td><td> TAAACAAGCT</td><td> 1260</td>

<td> TATGAAGAAA</td><td> TTAGCAAGGA</td><td> GCATTTGGCT</td><td> GTATATAAGA</td><td> TACAAATGTG</td><td> TAAAAGTGTT</td><td> 1320</td>

<td> AAAGCTCCAG</td><td> GAATATGTAT</td><td> TGATGTTGAT</td><td> AATGAAGATT</td><td> TGTTCTTTAT</td><td> AGCTGATAAA</td><td> 1380</td>

<td> AATAGTTTTT</td><td> CAGATGATTT</td><td> ATCTAAAAAC</td><td> GAAAGAATAG</td><td> AATATAATAC</td><td> ACAGAGTAAT</td><td> 1440</td>

<td> TATATAGAAA</td><td> ATGACTTCCC</td><td> TATAAATGAA</td><td> TTAATTTTAG</td><td> ATACTGATTT</td><td> AATAAGTAAA</td><td> 1500</td>

<td> ATAGAATTAC</td><td> CAAGTGAAAA</td><td> TACAGAATCA</td><td> CTTACTGATT</td><td> TTAATGTAGA</td><td> TGTTCCAGTA</td><td> 1560</td>

<td> TATGAAAAAC</td><td> AACCCGCTAT</td><td> AAAAAAAATT</td><td> TTTACAGATG</td><td> AAAATACCAT</td><td> CTTTCAATAT</td><td> 1620</td>

<td> TTATACTCTC</td><td> AGACATTTCC</td><td> TCTAGATATA</td><td> AGAGATATAA</td><td> GTTTAACATC</td><td> TTCATTTGAT</td><td> 1680</td>

<td> GATGCATTAT</td><td> TATTTTCTAA</td><td> CAAAGTTTAT</td><td> TCATTTTTTT</td><td> CTATGGATTA</td><td> TATTAAAACT</td><td> 1740</td>

<td> GCTAATAAAG</td><td> TGGTAGAAGC</td><td> AGGATTATTT</td><td> GCAGGTTGGG</td><td> TGAAACAGAT</td><td> AGTAAATGAT</td><td> 1800</td>

<td> TTTGTAATCG</td><td> AAGCTAATAA</td><td> AAGCAATACT</td><td> ATGGATAAAA</td><td> TTGCAGATAT</td><td> ATCTCTAATT</td><td> 1860</td>

<td> GTTCCTTATA</td><td> TAGGATTAGC</td><td> TTTAAATGTA</td><td> GGAAATGAAA</td><td> CAGCTAAAGG</td><td> AAATTTTGAA</td><td> 1920</td>

<td> AATGCTTTTG</td><td> AGATTGCAGG</td><td> AGCCAGTATT</td><td> CTACTAGAAT</td><td> TTATACCAGA</td><td> ACTTTTAATA</td><td> 1980</td>

<td> CCTGTAGTTG</td><td> GAGCCTTTTT</td><td> ATTAGAATCA</td><td> TATATTGACA</td><td> ATAAAAATAA</td><td> AATTATTAAA</td><td> 2040</td>

<td> ACAATAGATA</td><td> ATGCTTTAAC</td><td> TAAAAGAAAT</td><td> GAAAAATGGA</td><td> GTGATATGTA</td><td> CGGATTAATA</td><td> 2100</td>

<td> GTAGCGCAAT</td><td> GGCTCTCAAC</td><td> AGTTAATACT</td><td> CAATTTTATA</td><td> CAATAAAAGA</td><td> GGGAATGTAT</td><td> 2160</td>

<td> AAGGCTTTAA</td><td> ATTATCAAGC</td><td> ACAAGCATTG</td><td> GAAGAAATAA</td><td> TAAAATACAG</td><td> ATATAATATA</td><td> 2220</td>

<td> TATTCTGAAA</td><td> AAGAAAAGTC</td><td> AAATATTAAC</td><td> ATCGATTTTA</td><td> ATGATATAAA</td><td> TTCTAAACTT</td><td> 2280</td>

<td> AATGAGGGTA</td><td> TTAACCAAGC</td><td> TATAGATAAT</td><td> ATAAATAATT</td><td> TTATAAATGG</td><td> ATGTTCTGTA</td><td> 2340</td>

<td> TCATATTTAA</td><td> TGAAAAAAAT</td><td> GATTCCATTA</td><td> GCTGTAGAAA</td><td> AATTACTAGA</td><td> CTTTGATAAT</td><td> 2400</td>

<td> ACTCTCAAAA</td><td> AAAATTTGTT</td><td> AAATTATATA</td><td> GATGAAAATA</td><td> AATTATATTT</td><td> GATTGGAAGT</td><td> 2460</td>

<td> GCAGAATATG</td><td> AAAAATCAAA</td><td> AGTAAATAAA</td><td> TACTTGAAAA</td><td> CCATTATGCC</td><td> GTTTGATCTT</td><td> 2520</td>

<td> TCAATATATA</td><td> CCAATGATAC</td><td> AATACTAATA</td><td> GAAATGTTTA</td><td> ΑΤΑΑΑΤΑΤΆΑ</td><td> TAGC</td><td> 2574</td>

(2) INFORMATION FOR SEQ ID NO: 28:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2574 base pairs (B) TYPE: nucleic acid -- (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28:

ATGCCAGTTA CAATAAATAA TTTTAATTAT AATGATCCTA TTGATAATAA TAATATTATT60

ATGATGGAGC CTCCATTTGC GAGAGGTACG GGGAGATATT ATAAAGCTTT TAAAATCACA120

GATCGTATTT GGATAATACC GGAAAGATAT ACTTTTGGAT ATAAACCTGA GGATTTTAAT180

AAAAGTTCCG GTATTTTTAA TAGAGATGTT TGTGAATATT ATGATCCAGA TTACTTAAAT240

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- 113 ACTAATGATA AAAAGAATAT ATTTTTACAA ACAATGATCA AGTTATTTAA TAGAATCAAA 300 TCAAAACCAT TGGGTGAAAA GTTATTAGAG ATGATTATAA ATGGTATACC TTATCTTGGA 360 GATAGACGTG TTCCACTCGA AGAGTTTAAC ACAAACATTG CTAGTGTAAC TGTTAATAAA 420 TTAATCAGTA ATCCAGGAGA AGTGGAGCGA AAAAAAGGTA TTTTCGCAAA TTTAATAATA 4 80 TTTGGACCTG GGCCAGTTTT AAATGAAAAT GAGACTATAG ATATAGGTAT ACAAAATCAT 540 TTTGCATCAA GGGAAGGCTT CGGGGGTATA ATGCAAATGA AGTTTTGCCC AGAATATGTA 600 AGCGTATTTA ATAATGTTCA AGAAAACAAA GGCGCAAGTA TATTTAATAG ACGTGGATAT 660 TTTTCAGATC CAGCCTTGAT ATTAATGCAT GAACTCATCC ACGTCCTCCA CGGTCTCTAC 720 GGTATCAAAG TAGACGACCT CCCGATCGTC CCGAACGAAA AAAAATTCTT CATGCAGAGC 780 ACCGACGCAA TCCAGGCAGA AGAACTCTAC ACCTTCGGTG GTCAGGACCC GAGCATCATC 840 ACCCCGAGCA CCGACAAAAG CATCTACGAC AAAGTCCTCC AGAACTTCCG TGGTATCGTC 900 GACCGTCTCA ACAAAGTCCT CGTCTGCATC AGCGACCCGA ACATCAACAT CAACATCTAC 960 AAAAACAAAT TCAAAGACAA ATACAAATTC GTCGAAGACA GCGAAGGTAA ATACAGCATC 1020 GACGTCGAGA GCTTCGACAA ACTCTACAAA AGCCTCATGT TCGGTTTCAC CGAAACCAAC 1080 ATCGCAGAAA ACTACAAAAT CAAAACCCGT GCAAGCTACT TCAGCGACAG CCTCCCGCCG 1140 GTCAAAATCA AAAACCTCCT CGACAACGAA ATCTACACCA TCGAAGAAGG TTTCAACATC 1200 AGCGACAAAG ACATGGAAAA AGAATACCGT GGTCAGAACA AAGCAATCAA CAAACAAGCT 1260 TACGAAGAAA TCAGCAAAGA ACACCTCGCA GTCTACAAAA TCCAGATGTG CAAAAGCGTC 1320 AAAGCACCGG GTATCTGCAT CGACGTTGAC AACGAAGACC TCTTCTTCAT CGCAGACAAA 1380 AACAGCTTCA GCGACGACCT CAGCAAAAAC GAACGTATCG AATACAACAC CCAGAGCAAC 1440 TACATCGAAA ACGACTTCCC GATCAACGAA CTCATCCTCG ACACCGACCT CATCAGCAAA 1500 ATCGAACTCC CGAGCGAAAA CACCGAAAGC CTCACCGACT TCAACGTTGA CGTCCCGGTC 1560 TACGAAAAAC AGCCGGCAAT CAAAAAAATC TTCACCGACG AAAACACCAT CTTCCAGTAC 1620 CTCTACAGCC AGACCTTCCC GCTAGATATA AGAGATATAA GTTTAACATC TTCATTTGAT 1680 GATGCATTAT TATTTTCTAA CAAAGTTTAT TCATTTTTTT CTATGGATTA TATTAAAACT 1740 GCTAATAAAG TGGTAGAAGC AGGATTATTT GCAGGTTGGG TGAAACAGAT AGTAAATGAT 1800 TTTGTAATCG AAGCTAATAA AAGCAATACT ATGGATAAAA TTGCAGATAT ATCTCTAATT 1860 GTTCCTTATA TAGGATTAGC TTTAAATGTA GGAAATGAAA CAGCTAAAGG AAATTTTGAA 1920 AATGCTTTTG AGATTGCAGG AGCCAGTATT CTACTAGAAT TTATACCAGA ACTTTTAATA 1980 CCTGTAGTTG GAGCCTTTTT ATTAGAATCA TATATTGACA ATAAAAATAA AATTATTAAA 2 040 ACAATAGATA ATGCTTTAAC TAAAAGAAAT GAAAAATGGA GTGATATGTA CGGATTAATA 2100 GTAGCGCAAT GGCTCTCAAC AGTTAATACT CAATTTTATA CAATAAAAGA GGGAATGTAT 2160 AAGGCTTTAA ATTATCAAGC ACAAGCATTG GAAGAAATAA TAAAATACAG ATATAATATA 2220 TATTCTGAAA AAGAAAAGTC AAATATTAAC ATCGATTTTA ATGATATAAA TTCTAAACTT 2280

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AATGAGGGTA TTAACCAAGC TATAGATAAT

TCATATTTAA TGAAAAAAAT GATTCCATTA

ACTCTCAAAA AAAATTTGTT AAATTATATA

GCAGAATATG AAAAATCAAA AGTAAATAAA

TCAATATATA CCAATGATAC AATACTAATA

- 1 14 ATAAATAATT TTATAAATGG ATGTTCTGTA GCTGTAGAAA AATTACTAGA CTTTGATAAT GATGAAAATA AATTATATTT GATTGGAAGT TACTTGAAAA CCATTATGCC GTTTGATCTT

GAAATGTTTA ATAAATATAA TAGC

2340

2400

2460

2520

2574

Contents714

17 sheets

Sheet 1
Sheet 2
Sheet 3
Sheet 4
Sheet 5
Sheet 6
Sheet 7
Sheet 8
Sheet 9
Sheet 10
Sheet 11
Sheet 12
Sheet 13
Sheet 14
Sheet 15
Sheet 16
Sheet 17

Every citation, both ways

Citations
Document	Relation	Office	Cited during
WO9612802A1	Cites	World Intellectual Property Organization (WIPO)	Search report

49 members in 11 offices

This recordMember, by publication datePriority claim1 more office is in the list below

Priority claims12

Priority claims
Document	Office	Kind	Date
9617671	United Kingdom	A	1996-08-23
9617671	United Kingdom	A	1996-08-23
9625996	United Kingdom	A	1996-12-13
9625996	United Kingdom	A	1996-12-13
9702273	United Kingdom	W	1997-08-22
9702273	United Kingdom	W	1997-08-22
9617671	–	–	–
9625996	–	–	–
GB19960017671	–	–	–
GB19960025996	–	–	–
PCTGB9702273	–	–	–
WO1997GB02273	–	–	–

Members49

Family members, oldest first
Document	Office	Kind
GB9617671D0	United Kingdom	D0	1996-10-02
GB9625996D0	United Kingdom	D0	1997-01-29
CA2264191A1	Canada	A1	1998-02-26
WO9807864A1	World Intellectual Property Organization (WIPO)	A1	1998-02-26
AU4389597A	Australia	A	1998-03-06
ZA977541B	South Africa	B	1998-05-27
EP0939818A1	European Patent Office (EPO)	A1	1999-09-08
AU723397B2This record	Australia	B2	2000-08-24
JP2001502890A	Japan	A	2001-03-06
US2002044950A1	United States of America	A1	2002-04-18
US6461617B1	United States of America	B1	2002-10-08
US2003166238A1	United States of America	A1	2003-09-04
CA2498502A1	Canada	A1	2004-03-25
WO2004024909A2	World Intellectual Property Organization (WIPO)	A2	2004-03-25
AU2003267559A1	Australia	A1	2004-04-30
WO2004024909A3	World Intellectual Property Organization (WIPO)	A3	2004-07-08
EP0939818B1	European Patent Office (EPO)	B1	2005-04-27
AT294238T	Austria	T	2005-05-15
ATE294238T1	Austria	T1	2005-05-15
DE69733146D1	Germany	D1	2005-06-02
EP1537207A2	European Patent Office (EPO)	A2	2005-06-08
DK0939818T3	Denmark	T3	2005-08-29
US2005244435A1	United States of America	A1	2005-11-03
JP2005537809A	Japan	A	2005-12-15
DE69733146T2	Germany	T2	2006-03-09
US2006110410A1	United States of America	A1	2006-05-25
US7192596B2	United States of America	B2	2007-03-20
US2007184070A1	United States of America	A1	2007-08-09
US2007248626A1	United States of America	A1	2007-10-25
AU2003267559B2	Australia	B2	2008-08-14
JP4273250B2	Japan	B2	2009-06-03
US2009148888A1	United States of America	A1	2009-06-11
US2009246827A1	United States of America	A1	2009-10-01
US2009274708A1	United States of America	A1	2009-11-05
US2010022751A1	United States of America	A1	2010-01-28
US7674470B2	United States of America	B2	2010-03-09
EP2280066A2	European Patent Office (EPO)	A2	2011-02-02
US2011028691A1	United States of America	A1	2011-02-03
US7897158B2	United States of America	B2	2011-03-01
JP2011045377A	Japan	A	2011-03-10
EP2280066A3	European Patent Office (EPO)	A3	2011-05-11
US8012479B2	United States of America	B2	2011-09-06
US8012491B2	United States of America	B2	2011-09-06
US8017134B2	United States of America	B2	2011-09-13
CA2264191C	Canada	C	2012-01-17
US8454976B2	United States of America	B2	2013-06-04
CA2498502C	Canada	C	2014-02-11
JP5570936B2	Japan	B2	2014-08-13
US9006395B2	United States of America	B2	2015-04-14

1 legal event, as the office reported them to INPADOC

Events

Legal events, newest first
	Event	Code
2000-12-21	Letters patent sealed or granted (standard patent)GrantedFGA	FGA

Numbers

Publication, DOCDB: 723397
Publication, EPODOC: AU723397B
Application: 4389597
Application, DOCDB: 4389597
Application, EPODOC: AU19970043895

Titles

English: Recombinant toxin fragments

Classification

CPC, 23: C12N15/62
A61K38/00
A61K39/00
A61K39/08
A61K2039/505
A61K2039/53
A61K2039/627
C07K14/33
C07K14/475
C07K14/65
C07K2319/00
C07K2319/20
C07K2319/23
C07K2319/50
C07K2319/55
C07K2319/705
C07K2319/75
C12N9/52
Y10S530/825
A61K47/6415
A61K47/646
A61P31/04
Y02A50/30
Filled: the office’s first classification
IPC, 22: C12N15 09
A61K
A61K38 00
A61K38 16
A61K39 00
A61K39 08
A61K47 48
A61K48 00
A61P31 04
C07K14 33
C07K14 65
C07K19 00
C12N1 19
C12N1 21
C12N5 10
C12N9 52
C12N15 31
C12N15 62
C12P21 02
C12P21 04
C12R1 145
C12R1 19

Abstract

Term

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RECOMBINANT TOXIN FRAGMENTS

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- 29 SEQUENCE LISTING (1) GENERAL INFORMATION:

ATG CAG TTC GTG AAC AAG CAG TTC AAC TAT AAG GAC CCT GTA AAC GGT48

GTT GAC ATT GCC TAC ATC AAA ATT CCA AAC GCC GGC CAG ATG CAG CCG96

GTG AAG GCT TTC AAG ATT CAT AAC AAA ATC TGG GTT ATT CCG GAA CGC144

GAT ACA TTT ACG AAC CCG GAA GAA GGA GAC TTG AAC CCG CCG CCG GAA192

GCA AAG CAG GTG CCA GTT TCA TAC TAC GAT TCA ACC TAT CTG AGC ACA24 0

GAC AAC GAG AAG GAT AAC TAC CTG AAG GGA GTG ACC AAA TTA TTC GAG288

CGT ATT TAT TCC ACT GAC CTG GGC CGT ATG CTG CTG ACC TCA ATC GTC336

CGC GGA ATC CCA TTT TGG GGT GGC AGT ACC ATT GAC ACG GAG TTG AAG384

GTT ATT GAC ACT AAC TGC ATT AAC GTG ATC CAA CCA GAC GGT AGC TAC432

AGA TCT GAA GAA CTT AAC CTC GTA ATC ATC GGG CCC TCC GCG GAC ATT480

CGT AAC GGT TAC GGC TCT ACT CAG TAC ATT CGT TTC AGC CCA GAC TTC576

ACG TTC GGT TTC GAG GAG AGC CTG GAG GTT GAT ACC AAC CCG CTG TTG624

GGT GCA GGC AAG TTC GCA ACT GAT CCA GCG GTG ACC CTG GCA CAC GAG672

CTG ATC CAC GCC GGT CAT CGT CTG TAT GGC ATT GCG ATT AAC CCG AAC720

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- 31 CGC GTG TTC AAG GTT AAC ACC AAC GCC TAC TAC GAG ATG AGT GGT TTA768

GAA GTA AGC TTC GAG GAA CTG CGC ACG TTC GGT GGC CAT GAT GCG AAG816

TTT ATC GAC AGC TTG CAG GAG AAC GAG TTC CGT CTG TAC TAC TAC AAC864

AAG TTT AAA GAT ATT GCA AGT ACA CTG AAC AAG GCT AAG TCC ATT GTG912

GGT ACC ACT GCT TCA TTA CAG TAT ATG AAA AAT GTT TTT AAA GAG AAA960

TAT CTC CTA TCT GAA GAT ACA TCT GGA AAA TTT TCG GTA GAT. AAA TTA1008

AAA TTT GAT AAG TTA TAC AAA ATG TTA ACA GAG ATT TAC ACA GAG GAT1056

AAT TTT GTT AAG TTT TTT AAA GTA CTT AAC AGA AAA ACA TAT TTG AAT1104

TTT GAT AAA GCC GTA TTT AAG ATA AAT ATA GTA CCT AAG GTA AAT TAC1152

ACA ATA TAT GAT GGA TTT AAT TTA AGA AAT ACA AAT TTA GCA GCA AAC1200

TTT AAT GGT CAA AAT ACA GAA ATT AAT AAT ATG AAT TTT ACT AAA CTA124 8

AAA AAT TTT ACT GGA TTG TTT GAA TTT TAT AAG TTG CTA TGT GTA AGA1296

GGG ATA ATA ACT TCT AAA ACT AAA TCA TTA GAT AAA GGA TAC AAT AAG1344

GCA TTA AAT GAT TTA TGT ATC AAA GTT AAT AAT TGG GAC TTG TTT TTT1392

AGT CCT TCA GAA GAT AAT TTT ACT AAT GAT CTA AAT AAA GGA GAA GAA1440

GAT TTA ATA CAA CAA TAT TAT TTA ACC TTT AAT TTT GAT AAT GAA CCT1536