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filingDate 2002-06-24-04:00^^<http://www.w3.org/2001/XMLSchema#date>
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publicationDate 2007-04-18-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber EP-1411131-B1
titleOfInvention Process for producing template dna and process for producing protein in cell-free protein synthesis system with the use of the same
abstract Manufacturing template DNA for protein synthesis comprising PCR amplifying: (a) a first double-stranded DNA (dsDNA) fragment (D1) which encodes protein (fragment); and (b) contacting D1 with second and third dsDNA fragments (D2, D3) which overlap 5' and 3' ends of D1, respectively, where sense and antisense primers which anneal to 5' and 3' ends of D2 and D3, respectively, are used to initiate PCR, is new. Manufacturing (M1) template DNA for protein synthesis comprises PCR amplifying: (a) a first double-stranded DNA (dsDNA) fragment (D1) which encodes protein (fragment); and (b) contacting D1 with second and third dsDNA fragments (D2, D3) which overlap 5' and 3' ends of D1, respectively, where sense and antisense primers which anneal to 5' and 3' ends of D2 and D3, respectively, are used to initiate PCR amplification of the linear dsDNA, where D2 includes transcription and translation regulatory sequences and the amount of D2 and D3 in the PCR solution is respectively 5-2500 pmol/l. An independent claim is included for a cell-free protein synthesis method which involves use of a template DNA produced by M1.
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