发明公开
EP1358322A2 A METHOD OF INCREASING COMPLEMENTARITY IN A HETERODUPLEX POLYNUCLEOTIDE
有权
一种用于在异源核苷酸 - 增加互补
- 专利标题: A METHOD OF INCREASING COMPLEMENTARITY IN A HETERODUPLEX POLYNUCLEOTIDE
- 专利标题(中): 一种用于在异源核苷酸 - 增加互补
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申请号: EP02741635.3申请日: 2002-02-01
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公开(公告)号: EP1358322A2公开(公告)日: 2003-11-05
- 发明人: PADGETT, Hal, S. , FITZMAURICE, Wayne, P. , LINDO, John, A.
- 申请人: Large Scale Biology Corporation
- 申请人地址: Suite 1000,3333 Vaca Valley Parkway Vacaville, CA 95688 US
- 专利权人: Large Scale Biology Corporation
- 当前专利权人: Large Scale Biology Corporation
- 当前专利权人地址: Suite 1000,3333 Vaca Valley Parkway Vacaville, CA 95688 US
- 代理机构: Brasnett, Adrian Hugh
- 优先权: US266386P 20010202; US268785P 20010214
- 国际公布: WO02079468 20021010
- 主分类号: C12N15/10
- IPC分类号: C12N15/10
摘要:
We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3' to 5' exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotide will necessarily have all mismatches resolved to complementary. The resulting polynucleotide is optionally ligated. Several variant polynucleotide result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. The parent polynucleotides need not be cleaved into fragments prior to annealing heterologous strands. Therefore, no reassembly is required.
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