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1. 发明申请

WO02079468A8 A METHOD OF INCREASING COMPLEMENTARITY IN A HETERODUPLEX POLYNUCLEOTIDE 审中-公开
标题翻译：一种提高二叉多项式多项式中的互补性的方法
公开(公告)号：WO02079468A8
公开(公告)日：2003-12-04
申请号：PCT/US0203055
申请日：2002-02-01
申请人： LARGE SCALE BIOLOGY CORP , PADGETT HAL S , FITZMAURICE WAYNE P , LINDO JOHN A
发明人： PADGETT HAL S , FITZMAURICE WAYNE P , LINDO JOHN A
IPC分类号： C12N15/09 , C12N15/10 , C12P21/00
CPC分类号： C12N15/102 , C12N15/1027
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.
摘要翻译：我们在此描述增加异源双链多核苷酸序列中互补性的体外方法。该方法使用相反链退火形成具有错配的多核苷酸双链体。将异源双链体多核苷酸与有效量的具有链切割活性，3'至5'外切核酸酶活性和聚合酶活性的酶组合，并且允许足够的时间以在异源双链体内增加互补百分比。不是所有的异源双链多核苷酸都必须将所有错配解析为互补。所得多核苷酸任选连接。几个变体多核苷酸结果在其中任一条相反链在另一条链中模板化重新编码的位点，异源双链体多核苷酸序列的所得百分比互补性增加。亲本多核苷酸在退火异源链之前不需要被切割成片段。因此，不需要重新组装。

2. 发明申请

WO2002079468A3 A METHOD OF INCREASING COMPLEMENTARITY IN A HETERODUPLEX POLYNUCLEOTIDE 审中-公开
公开(公告)号：WO2002079468A3
公开(公告)日：2002-10-10
申请号：PCT/US2002/003055
申请日：2002-02-01
申请人： LARGE SCALE BIOLOGY CORPORATION , PADGETT, Hal, S. , FITZMAURICE, Wayne, P. , LINDO, John, A.
发明人： PADGETT, Hal, S. , FITZMAURICE, Wayne, P. , LINDO, John, A.
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.

3. 发明申请

WO2002079468A2 A METHOD OF INCREASING COMPLEMENTARITY IN A HETERODUPLEX 审中-公开
标题翻译：一种在异位中增加补充的方法
公开(公告)号：WO2002079468A2
公开(公告)日：2002-10-10
申请号：PCT/US2002/003055
申请日：2002-02-01
申请人： LARGE SCALE BIOLOGY CORPORATION , PADGETT, Hal, S. , FITZMAURICE, Wayne, P. , LINDO, John, A.
发明人： PADGETT, Hal, S. , FITZMAURICE, Wayne, P. , LINDO, John, A.
IPC分类号： C12N15/10
CPC分类号： C12N15/102 , C12N15/1027
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.
摘要翻译：我们在这里描述了增加异源双链多核苷酸序列中的互补性的体外方法。该方法使用相反链的退火以形成具有错配的多核苷酸双链体。将异源双链多核苷酸与有效量的具有链切割活性，3'至5'核酸外切酶活性和聚合酶活性的酶组合，并且允许足够的时间在异源双链体内增加互补百分比。并非所有的异源双链体多核苷酸都必然具有解决互补的所有错配。任选地连接得到的多核苷酸。产生几种变体多核苷酸。在相对链中的任一个在另一条链中模板记录的位点，异源双链多核苷酸序列的所得百分比互补性增加。在退火异源链之前，母体多核苷酸不需要切割成片段。因此，不需要重新组装。

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