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    Methods and compositions for generating recombinant nucleic acid molecules
    1.
    发明授权
    Methods and compositions for generating recombinant nucleic acid molecules 有权
    用于产生重组核酸分子的方法和组合物

    公开(公告)号：US08207318B2

    公开(公告)日：2012-06-26

    申请号：US12112649

    申请日：2008-04-30

    申请人： James Fan ,  John Carrino ,  Jonathan Chesnut ,  Knut Madden ,  Martin Gleeson ,  Robert Bennett

    发明人： James Fan ,  John Carrino ,  Jonathan Chesnut ,  Knut Madden ,  Martin Gleeson ,  Robert Bennett

    IPC分类号： C07H21/02 ,  C12Q1/68

    CPC分类号： C12N15/10 ,  C12N9/90 ,  C12N15/111 ,  C12N15/64 ,  C12N15/66 ,  C12N2310/14 ,  C12N2330/30 ,  C12P19/34 ,  C12Q1/6855 ,  C12Y599/01002 ,  C12Q2521/519

    摘要： A method of generating a double stranded (ds) recombinant nucleic acid molecule covalently linked in both strands by contacting two or more ds nucleotide sequences with a topoisomerase under conditions such that both termini of at least one end of a first ds nucleotide sequence are covalently linked by the topoisomerase to both termini of at least one end of a second ds nucleotide sequence is provided. Also provided is a method for generating a ds recombinant nucleic acid molecule covalently linked in one strand, by contacting two or more ds nucleotide sequences with a type IA topoisomerase under conditions such that one strand, but not both strands, of one or both ends of a first ds nucleotide sequence are covalently linked by the topoisomerase. Compositions for performing such methods, and compositions generated from such methods also are provided, as are kits containing components useful for conveniently practicing the methods.

    摘要翻译： 通过使两个或多个ds核苷酸序列与拓扑异构酶接触的条件使得在第一个ds核苷酸序列的至少一个末端的两个末端共价连接的情况下，产生双链（ds）重组核酸分子共价连接的双链（ds）重组核酸分子的方法 提供了拓扑异构酶至第二个ds核苷酸序列的至少一个末端的两个末端。 还提供了一种用于产生共价连接在一条链中的ds重组核酸分子的方法，其通过使两个或更多个ds核苷酸序列与IA型拓扑异构酶接触，使得一个或两个末端的一条链但不是两条链 第一个ds核苷酸序列由拓扑异构酶共价连接。 还提供了用于进行这些方法的组合物，以及由这些方法产生的组合物，以及包含用于方便地实施该方法的组分的试剂盒。

    COMPOSITIONS AND METHODS FOR RAPIDLY GENERATING RECOMBINANT NUCLEIC ACID MOLECULES
    2.
    发明申请
    COMPOSITIONS AND METHODS FOR RAPIDLY GENERATING RECOMBINANT NUCLEIC ACID MOLECULES 有权
    用于快速产生重组核酸分子的组合物和方法

    公开(公告)号：US20060029935A1

    公开(公告)日：2006-02-09

    申请号：US10014128

    申请日：2001-12-07

    申请人： John Carrino ,  James Fan ,  Robert Bennett ,  Jonathan Chesnut ,  Martin Gleeson ,  Knut Madden

    发明人： John Carrino ,  James Fan ,  Robert Bennett ,  Jonathan Chesnut ,  Martin Gleeson ,  Knut Madden

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12P19/34 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/66 ,  C12Q1/6855 ,  C12Q2521/519

    摘要： A method of generating a double stranded (ds) recombinant nucleic acid molecule covalently linked in both strands by contacting two or more ds nucleotide sequences with a topoisomerase under conditions such that both termini of at least one end of a first ds nucleotide sequence are covalently linked by the topoisomerase to both termini of at least one end of a second ds nucleotide sequence is provided. Also provided is a method for generating a ds recombinant nucleic acid molecule covalently linked in one strand, by contacting two or more ds nucleotide sequences with a type IA topoisomerase under conditions such that one strand, but not both strands, of one or both ends of a first ds nucleotide sequence are covalently linked by the topoisomerase. Compositions for performing such methods, and compositions generated from such methods also are provided, as are kits containing components useful for conveniently practicing the methods.

    摘要翻译： 通过使两个或多个ds核苷酸序列与拓扑异构酶接触的条件使得在第一个ds核苷酸序列的至少一个末端的两个末端共价连接的情况下，产生双链（ds）重组核酸分子共价连接的双链（ds）重组核酸分子的方法 提供了拓扑异构酶至第二个ds核苷酸序列的至少一个末端的两个末端。 还提供了一种用于产生共价连接在一条链中的ds重组核酸分子的方法，其通过使两个或更多个ds核苷酸序列与IA型拓扑异构酶接触，使得一个或两个末端的一条链但不是两条链 第一个ds核苷酸序列由拓扑异构酶共价连接。 还提供了用于进行这些方法的组合物，以及由这些方法产生的组合物，以及包含用于方便地实施该方法的组分的试剂盒。

    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLE RECOGNITION SITES
    4.
    发明申请
    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLE RECOGNITION SITES 审中-公开
    使用多个识别位点合成核酸分子的方法和组合物

    公开(公告)号：US20070196838A1

    公开(公告)日：2007-08-23

    申请号：US11612957

    申请日：2006-12-19

    申请人： Jonathan Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James Hartley ,  Devon Byrd ,  Gray Temple

    发明人： Jonathan Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James Hartley ,  Devon Byrd ,  Gray Temple

    IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12P21/06 ,  C12N9/22

    CPC分类号： C12N9/90 ,  C12P19/34

    摘要： The present invention provides compositions and methods for recombinational cloning. The compositions vectors having multiple recombination sites and/or multiple topoisomerase recognition sities. The methods premit the simultaeous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according to the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

    摘要翻译： 本发明提供了用于重组克隆的组合物和方法。 具有多个重组位点和/或多个拓扑异构酶识别性质的组合物载体。 该方法预先提出了两种或多种不同核酸分子的同源克隆。 在一些实施方案中，分子融合在一起，而在其它实施方案中，分子被插入载体中的不同位点。 本发明还通常提供通过重组连接或连接许多可以相同或不同的分子和/或化合物（例如，化合物，药物，蛋白质或肽，脂质，核酸，碳水化合物等）。 本发明还提供了包含本发明的核酸分子或根据本发明的方法制备的宿主细胞，还提供了包含本发明的组合物，宿主细胞和核酸分子的试剂盒，其可用于合成核酸分子 根据本发明的方法。