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1. 发明授权

US08030066B2 Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites 有权
标题翻译：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：US08030066B2
公开(公告)日：2011-10-04
申请号：US11612445
申请日：2006-12-18
申请人： Jonathan D. Chesnut , John Carrino , Louis Leong , Knut Madden , Martin Gleeson , James Fan , Michael Brasch , David Cheo , James L. Hartley , Devon R. Byrd , Gray F. Temple
发明人： Jonathan D. Chesnut , John Carrino , Louis Leong , Knut Madden , Martin Gleeson , James Fan , Michael Brasch , David Cheo , James L. Hartley , Devon R. Byrd , Gray F. Temple
IPC分类号： C12N15/00
CPC分类号： C12N15/66 , C12N9/90 , C12N15/10 , C12N15/64 , C12N15/902 , C12N2800/108 , C12N2800/30 , C12N2800/70 , C12N2840/20 , C12P19/34 , C12Y599/01
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.
摘要翻译：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。

2. 发明申请

US20090186387A1 RECOMBINATIONAL CLONING USING ENGINEERED RECOMBINATION SITES 审中-公开
标题翻译：使用工程改造站点进行重组克隆
公开(公告)号：US20090186387A1
公开(公告)日：2009-07-23
申请号：US12247861
申请日：2008-10-08
申请人： James L. Hartley , Michael Brasch
发明人： James L. Hartley , Michael Brasch
IPC分类号： C12P19/34
CPC分类号： C12P19/34 , C12N9/00 , C12N15/10 , C12N15/64 , C12N15/66
摘要： Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).
摘要翻译：通过在体外和体内使用核酸，载体和方法，使用工程化重组位点和重组蛋白来移动或交换DNA分子片段来提供重组克隆，以提供具有所需特征的嵌合DNA分子和 /或DNA片段。

3. 发明申请

US20090186385A1 RECOMBINATIONAL CLONING USING NUCLEIC ACIDS HAVING RECOMBINATION SITES 审中-公开
标题翻译：使用具有重组位点的核酸进行重组克隆
公开(公告)号：US20090186385A1
公开(公告)日：2009-07-23
申请号：US12207382
申请日：2008-09-09
申请人： James L. Hartley , Michael Brasch , Gary F. Temple , Donna K. Fox
发明人： James L. Hartley , Michael Brasch , Gary F. Temple , Donna K. Fox
IPC分类号： C12P19/34
CPC分类号： C12N15/64 , C12N9/00 , C12N15/10 , C12N15/66
摘要： Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).
摘要翻译：通过在体外和体内使用核酸，载体和方法，使用工程化重组位点和重组蛋白来移动或交换DNA分子片段来提供重组克隆，以提供具有所需特征的嵌合DNA分子和 /或DNA片段。

4. 发明申请

US20110033920A1 COMPOSITIONS AND METHODS FOR USE IN RECOMBINATIONAL CLONING OF NUCELIC ACIDS 有权
标题翻译：用于重组克隆核酸的组合物和方法
公开(公告)号：US20110033920A1
公开(公告)日：2011-02-10
申请号：US12649129
申请日：2009-12-29
申请人： James L. Hartley , Michael A. Brasch , Gary F. Temple , David Cheo
发明人： James L. Hartley , Michael A. Brasch , Gary F. Temple , David Cheo
IPC分类号： C12N15/63 , C07H21/04
CPC分类号： C12N15/11 , C12N15/10 , C12N15/63 , C12N2800/70
摘要： The present invention relates generally to compositions and methods for use in recombinational cloning of nucleic acid molecules. In particular, the invention relates to nucleic acid molecules encoding one or more recombination sites or portions thereof, to nucleic acid molecules comprising one or more of these recombination site nucleotide sequences and optionally comprising one or more additional physical or functional nucleotide sequences. The invention also relates to vectors comprising the nucleic acid molecules of the invention, to host cells comprising the vectors or nucleic acid molecules of the invention, to methods of producing polypeptides using the nucleic acid molecules of the invention, and to polypeptides encoded by these nucleic acid molecules or produced by the methods of the invention. The invention also relates to antibodies that bind to one or more polypeptides of the invention or epitopes thereof. The invention also relates to the use of these compositions in methods for recombinational cloning of nucleic acids, in vitro and in vivo, to provide chimeric DNA molecules that have particular characteristics and/or DNA segments.
摘要翻译：本发明一般涉及用于核酸分子重组克隆的组合物和方法。特别地，本发明涉及编码一个或多个重组位点或其部分的核酸分子与包含这些重组位点核苷酸序列中的一个或多个并且任选地包含一个或多个另外的物理或功能核苷酸序列的核酸分子。本发明还涉及包含本发明的核酸分子的载体，包含本发明的载体或核酸分子的宿主细胞，使用本发明的核酸分子产生多肽的方法，以及由这些核酸编码的多肽酸分子或通过本发明的方法制备。本发明还涉及结合本发明的一种或多种多肽或其表位的抗体。本发明还涉及这些组合物用于在体外和体内重组克隆核酸以提供具有特定特征和/或DNA片段的嵌合DNA分子的方法中的用途。

5. 发明授权

US07408049B2 Recombinational cloning using nucleic acids having recombination sites 有权
标题翻译：使用具有重组位点的核酸进行重组克隆
公开(公告)号：US07408049B2
公开(公告)日：2008-08-05
申请号：US10893235
申请日：2004-07-19
申请人： James L. Hartley , Michael A. Brasch , Gary F. Temple , Donna K. Fox
发明人： James L. Hartley , Michael A. Brasch , Gary F. Temple , Donna K. Fox
IPC分类号： C12N15/11
CPC分类号： C12N15/64 , C12N9/00 , C12N15/10 , C12N15/66
摘要： Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).
摘要翻译：通过在体外和体内使用核酸，载体和方法，使用工程化重组位点和重组蛋白来移动或交换DNA分子片段来提供重组克隆，以提供具有所需特征的嵌合DNA分子和 /或DNA片段。

6. 发明授权

US07223576B2 Recombinational cloning using engineered recombination sites 失效
标题翻译：使用工程重组位点的重组克隆
公开(公告)号：US07223576B2
公开(公告)日：2007-05-29
申请号：US10796868
申请日：2004-03-10
申请人： James L. Hartley , Michael A. Brasch
发明人： James L. Hartley , Michael A. Brasch
IPC分类号： C12N9/00 , C12N15/70
CPC分类号： C12N9/00 , C12N15/10 , C12N15/64 , C12N15/66
摘要： Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).
摘要翻译：通过在体外和体内使用核酸，载体和方法，使用工程化重组位点和重组蛋白来移动或交换DNA分子片段来提供重组克隆，以提供具有所需特征的嵌合DNA分子和 /或DNA片段。

7. 发明授权

US06518026B2 Process for controlling contamination of nucleic acid amplification reactions 有权
标题翻译：控制核酸扩增反应污染的方法
公开(公告)号：US06518026B2
公开(公告)日：2003-02-11
申请号：US09782516
申请日：2001-02-14
申请人： James L. Hartley
发明人： James L. Hartley
IPC分类号： C12Q168
CPC分类号： C12Q1/6848 , C12Q1/6853 , C12Q1/708 , Y10S435/81 , C12Q2525/117 , C12Q2521/531 , C12Q2531/113 , C12Q2525/186 , C12Q2531/137 , C12Q2521/301 , C12Q2537/137 , C12Q2525/101
摘要： This invention relates to a method of incorporating an exo-sample nucleotide into the amplified product strands resulting from a nucleic acid amplification process. Once the product strands have been obtained and analyzed (e.g., by hybridization, Southern blot, etc.), the exo-sample strands can be selectively destroyed by acting on the incorporated exo-sample nucleotide. Two embodiments are presented. In a first embodiment, the exo-sample nucleotide is incorporated by carrying out the amplification reaction in the presence of an excess of exo-sample nucleotide triphosphate. In a second embodiment, the exo-sample nucleotide is incorporated by carrying out the amplification reaction in the presence of an oligonucleotide which has, as part of its sequence, one or more exo-sample nucleotides.
摘要翻译：本发明涉及将外切样品核苷酸掺入由核酸扩增过程产生的扩增产物链中的方法。一旦获得并分析了产物链（例如通过杂交，Southern印迹等），可以通过作用于外加的外切核苷酸来选择性地破坏外部样品链。提供了两个实施方案。在第一个实施方案中，通过在过量样品中的三磷酸过多的存在下进行扩增反应来引入外切核苷酸。在第二个实施方案中，通过进行扩增反应来引入外切核苷酸作为其序列的一部分具有一个或多个外切核苷酸的寡核苷酸的存在。

8. 发明授权

US6143557A Recombination cloning using engineered recombination sites 有权
标题翻译：使用工程重组位点的重组克隆
公开(公告)号：US6143557A
公开(公告)日：2000-11-07
申请号：US233493
申请日：1999-01-20
申请人： James L. Hartley , Michael A. Brasch
发明人： James L. Hartley , Michael A. Brasch
IPC分类号： C12N9/00 , C12N15/10 , C12N15/64 , C12N15/66 , C12N15/63 , C07H21/04 , C12N15/68
CPC分类号： C12P19/34 , C12N15/10 , C12N15/64 , C12N15/66 , C12N9/00
摘要： Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).
摘要翻译：通过在体外和体内使用核酸，载体和方法，使用工程化重组位点和重组蛋白来移动或交换DNA分子片段来提供重组克隆，以提供具有所需特征的嵌合DNA分子和 /或DNA片段。

9. 发明授权

US5970782A Gradient filtration apparatus 失效
标题翻译：梯度过滤装置
公开(公告)号：US5970782A
公开(公告)日：1999-10-26
申请号：US79174
申请日：1998-05-15
申请人： James L. Hartley , Terry A. Landers
发明人： James L. Hartley , Terry A. Landers
IPC分类号： B01D61/14 , B01D61/18 , C12M1/12 , G01N15/02 , G01N15/08 , B07B1/50
CPC分类号： B01D61/147 , B01D61/18 , C12M29/04 , C12M41/36 , G01N15/0272
摘要： Particles such as bacteria may be enumerated from a liquid in an accurate and efficient manner through use of an apparatus including a filter sheet which is oriented transverse to the surface of the liquid being filtered. The filter sheet is so arranged so that, as the liquid is filtered, an increasing portion of the filter is above the surface of the liquid. As a result of the surface of the liquid dropping across the filter, a smaller fraction of the total volume of the liquid passes through the upper portion of the filter than through the lower portion of the filter. Since the number of bacteria trapped per unit area of the filter depends on the volume filtered through the area, a monotonically increasing gradient of density of filtered bacteria occurs from the top to the bottom of the filter.
摘要翻译：可以通过使用包括与被过滤的液体的表面横向定向的过滤片的设备，以精确和有效的方式从液体中列举诸如细菌的颗粒。过滤片被布置成使得当过滤液体时，过滤器的增加部分在液体表面之上。由于液体表面滴落在过滤器上，液体的总体积的较小部分通过过滤器的上部，而不是通过过滤器的下部。由于过滤器每单位面积捕获的细菌数量取决于通过该区域过滤的体积，所以从过滤器的顶部到底部发生单调递增的过滤细菌浓度梯度。

10. 发明申请

US20100267119A1 RECOMBINATIONAL CLONING USING NUCLEIC ACIDS HAVING RECOMBINATION SITES 审中-公开
公开(公告)号：US20100267119A1
公开(公告)日：2010-10-21
申请号：US12497832
申请日：2009-07-06
申请人： James L. Hartley , Michael A. Brasch , Gary F. Temple , Donna K. Fox
发明人： James L. Hartley , Michael A. Brasch , Gary F. Temple , Donna K. Fox
IPC分类号： C12N1/21 , C12N15/74
CPC分类号： C12N15/64 , C12N9/00 , C12N15/10 , C12N15/66
摘要： Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).

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