发明名称 |
Recombination of polynucleotide sequences using random or defined primers |
摘要 |
A method for in vitro mutagenesis and recombination of polynucleotide sequences based on polymerase-catalyzed extension of primer oligonucleotides is disclosed. The method involves priming template polynucleotide(s) with random-sequences or defined-sequence primers to generate a pool of short DNA fragments with a low level of point mutations. The DNA fragments are subjected to denaturization followed by annealing and further enzyme-catalyzed DNA polymerization. This procedure is repeated a sufficient number of times to produce full-length genes which comprise mutants of the original template polynucleotides. These genes can be further amplified by the polymerase chain reaction and cloned into a vector for expression of the encoded proteins.
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申请公布号 |
US6153410(A) |
申请公布日期 |
2000.11.28 |
申请号 |
US19970905359 |
申请日期 |
1997.08.04 |
申请人 |
CALIFORNIA INSTITUTE OF TECHNOLOGY |
发明人 |
ARNOLD, FRANCES H.;SHAO, ZHIXIN;AFFHOLTER, JOSEPH A.;ZHAO, HUIMIN H;GIVER, LORRAINE J. |
分类号 |
C12N15/09;C12N9/16;C12N9/56;C12N9/80;C12N15/10;C12Q1/68;(IPC1-7):C12N15/09;C12P19/34 |
主分类号 |
C12N15/09 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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