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11.发明授权公开(公告)号：US08338091B2
公开(公告)日：2012-12-25
申请号：US13401680
申请日：2012-02-21
申请人： Jonathan Chesnut , Miroslav Dudas , Adam Harris , Louis Leong , Knut Madden
发明人： Jonathan Chesnut , Miroslav Dudas , Adam Harris , Louis Leong , Knut Madden
IPC分类号： C12Q1/68
CPC分类号： C12P19/34 , C07H21/04 , C12N9/90 , C12N15/10 , C12N15/111 , C12N15/64 , C12N15/66 , C12N15/86 , C12N2310/111 , C12N2310/14 , C12N2330/30 , C12N2740/16043 , C12N2800/30 , C12N2800/70 , C12Y599/01
摘要： The present invention is in the fields of biotechnology and molecular biology. More particularly, the present invention relates to cloning or subcloning one or more nucleic acid molecules comprising one or more type IIs restriction enzyme recognition sites. The present invention also embodies cloning such nucleic acid molecules using recombinational cloning methods such as those employing recombination sites and recombination proteins. The present invention also relates to nucleic acid molecules (including RNA and iRNA), as well as proteins, expressed from host cells produced using the methods of the present invention.
摘要翻译： 本发明涉及生物技术和分子生物学领域。 更具体地说，本发明涉及克隆或亚克隆一个或多个包含一个或多个II型限制酶识别位点的核酸分子。 本发明还使用重组克隆方法（例如使用重组位点和重组蛋白的那些）来体现克隆这种核酸分子。 本发明还涉及由使用本发明的方法生产的宿主细胞表达的核酸分子（包括RNA和iRNA）以及蛋白质。
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12.发明授权公开(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
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。
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13.发明授权公开(公告)号：US07244560B2
公开(公告)日：2007-07-17
申请号：US10005876
申请日：2001-12-07
申请人： Jonathan D. Chestnut , John Carrino , Louis Leong , Knut Madden , Martin Gleeson , James Fan , Michael A. Brasch , David Cheo , James L. Hartley , Devon R.N. Byrd , Gary F. Temple
发明人： Jonathan D. Chestnut , John Carrino , Louis Leong , Knut Madden , Martin Gleeson , James Fan , Michael A. Brasch , David Cheo , James L. Hartley , Devon R.N. Byrd , Gary F. Temple
IPC分类号： C12Q1/68
摘要： The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous 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.
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14.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
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。
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公开(公告)号：US20060008817A1
公开(公告)日：2006-01-12
申请号：US10991803
申请日：2004-11-17
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
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.
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