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    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLERECOGNITION SITES
    1.
    发明申请
    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLERECOGNITION SITES 有权
    使用多重分解位点合成核酸分子的方法和组合物

    公开(公告)号：US20130323796A1

    公开(公告)日：2013-12-05

    申请号：US13892903

    申请日：2013-05-13

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Jonathan CHESNUT ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James Hartley ,  Devon Byrd ,  Gary Temple

    IPC: C12P19/34

    CPC classification number: C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

    Abstract: 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.

    Abstract translation: 本发明提供了用于重组克隆的组合物和方法。 该组合物包括具有多个重组裂解物和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 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数个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。

    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLERECOGNITION SITES
    4.
    发明申请
    METHODS AND COMPOSITIONS FOR SYNTHESIS OF NUCLEIC ACID MOLECULES USING MULTIPLERECOGNITION SITES 有权
    使用多重分解位点合成核酸分子的方法和组合物

    公开(公告)号：US20150225729A1

    公开(公告)日：2015-08-13

    申请号：US14578336

    申请日：2014-12-19

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Jonathan CHESNUT ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James Hartley ,  Devon Byrd ,  Gary Temple

    IPC: C12N15/66 ,  C12N9/90

    CPC classification number: C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

    Abstract: 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.

    Abstract translation: 本发明提供了用于重组克隆的组合物和方法。 组合物包括具有多个重组位点和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 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。

    Methods and compositions for synthesis of nucleic acid molecules using multiplerecognition sites
    5.
    发明授权
    Methods and compositions for synthesis of nucleic acid molecules using multiplerecognition sites 有权
    使用多重识别位点合成核酸分子的方法和组合物

    公开(公告)号：US08945884B2

    公开(公告)日：2015-02-03

    申请号：US13892903

    申请日：2013-05-13

    Applicant: Life Technologies Corporation

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

    IPC: C12N15/64 ,  C12P19/34 ,  C12N15/10 ,  C12N15/66 ,  C12N15/90

    CPC classification number: C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

    Abstract: 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.

    Abstract translation: 本发明提供了用于重组克隆的组合物和方法。 组合物包括具有多个重组位点和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 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。

    METHODS AND REAGENTS FOR MOLECULAR CLONING
    8.
    发明申请
    METHODS AND REAGENTS FOR MOLECULAR CLONING 审中-公开
    分子克隆的方法和试剂

    公开(公告)号：US20150093788A1

    公开(公告)日：2015-04-02

    申请号：US14505396

    申请日：2014-10-02

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Jonathan CHESNUT ,  Stewart Shuman ,  Knut Madden ,  John Heyman ,  Robert Bennett

    IPC: C12N15/66 ,  C12N15/64

    CPC classification number: C12N15/66 ,  C12N15/10 ,  C12N15/64

    Abstract: The present invention provides compositions, methods, and kits for covalently linking nucleic acid molecules. The methods include a strand invasion step, and the compositions and kits are useful for performing such methods. For example, a method of covalently linking double stranded (ds) nucleic acid molecules can include contacting a first ds nucleic acid molecule, which has a topoisomerase linked to a 3′ terminus of one end and has a single stranded 5′ overhang at the same end, with a second ds nucleic acid molecule having a blunt end, such that the 5′ overhang can hybridize to a complementary sequence of the blunt end of the second nucleic acid molecule, and the topoisomerase can covalently link the ds nucleic acid molecules. The methods are simpler and more efficient than previous methods for covalently linking nucleic acid sequences, and the compositions and kits facilitate practicing the methods, including methods of directionally linking two or more ds nucleic acid molecules.

    Abstract translation: 本发明提供用于共价连接核酸分子的组合物，方法和试剂盒。 所述方法包括线入侵步骤，组合物和试剂盒对于进行这些方法是有用的。 例如，共价连接双链（ds）核酸分子的方法可以包括使第一个ds核酸分子接触，该第一个ds核酸分子具有与一个末端的3'末端连接的拓扑异构酶并且具有相同的单链（5'）突出端 末端，具有平末端的第二ds核酸分子，使得5'突出端可以与第二核酸分子的平端的互补序列杂交，并且拓扑异构酶可以共价连接ds核酸分子。 该方法比先前用于共价连接核酸序列的方法更简单和更有效，并且组合物和试剂盒有助于实施该方法，包括定向连接两个或多个ds核酸分子的方法。

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