Process for producing template dna and process for producing protein in cell-free protein synthesis system with the use of the same

(57) Offer the efficient manufacture methods of mold DNA for revealing and refining summary subject protein. A solution means -- the 1st 2 chain DNA fragment that carries out the code of the protein -- this -- with the 2nd 2 chain DNA fragment including the arrangement which overlaps with 5' distal region of the 1st DNA fragment, this -- with the 3rd 2 chain DNA fragment including the arrangement which overlaps with 3' distal region of the 1st DNA fragment, this -- with the sense primer which carries out annealing to 5' distal region of the 2nd DNA fragment, and -- this -- in the method of contacting the antisense primer which carries out annealing to 3' distal region of the 3rd DNA fragment, and amplifying two straight-chain-shape chain DNA by polymerase chain reaction (PCR), this -- the arrangement whose 2nd DNA fragment controls transfer and translation of a gene is included, and the production method of mold DNA for protein composition with which concentration of the 2nd DNA fragment in this PCR solution and the 3rd DNA fragment is characterized by being 5*2500 pmol/L, respectively is offered.