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11.发明授权Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme 失效使用热稳定酶扩增，检测和/或克隆核酸序列的方法
公开(公告)号：US4965188A
公开(公告)日：1990-10-23
申请号：US63647
申请日：1987-06-17
CPC分类号： C12P19/34 , C12N9/1252 , C12Q1/6844
摘要： A process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers and extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence. The amplified sequence can be readily detected. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed.
摘要翻译： 用于扩增核酸或其混合物中包含的任何靶核酸序列的方法包括用摩尔过量的两个寡核苷酸引物处理所述核酸的分开的互补链，并用热稳定酶扩增引物以形成互补引物延伸产物，其起作用 作为合成所需核酸序列的模板。 可以容易地检测扩增的序列。 反应的步骤可以根据需要经常重复，并且涉及温度循环以进行杂交，促进酶的活性和所形成的杂交体的变性。
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公开(公告)号：US06514736B1
公开(公告)日：2003-02-04
申请号：US09704357
申请日：2000-11-01
IPC分类号： C12N912
CPC分类号： C12Q1/6858 , B01J19/0046 , B01J2219/0059 , B01J2219/00722 , B01L7/52 , C07K14/70539 , C07K14/805 , C12N9/1252 , C12N15/10 , C12P19/34 , C12Q1/6827 , C12Q1/6853 , C12Q1/686 , C40B40/06 , Y10S977/92 , C12Q2549/119 , C12Q2525/149 , C12Q2525/143 , C12Q2525/131
摘要： The present invention is directed to a process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof using a thermostable enzyme. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed.
摘要翻译： 本发明涉及使用热稳定性酶扩增核酸或其混合物中包含的任何靶核酸序列的方法。 该方法包括用摩尔过量的两种寡核苷酸引物处理核酸的分开的互补链，用热稳定酶扩增引物以形成互补引物延伸产物，其用作合成所需核酸序列的模板，并检测序列 放大 反应的步骤可以根据需要经常重复，并且涉及温度循环以进行杂交，促进酶的活性和所形成的杂交体的变性。
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13.发明授权
公开(公告)号：US6040166A
公开(公告)日：2000-03-21
申请号：US312729
申请日：1994-09-27
IPC分类号： B01J19/00 , B01L7/00 , C07K14/74 , C07K14/805 , C12N9/12 , C12N15/10 , C12P19/34 , C12Q1/68 , C40B40/06
CPC分类号： C12Q1/6858 , B01J19/0046 , B01L7/52 , C07K14/70539 , C07K14/805 , C12N15/10 , C12N9/1252 , C12P19/34 , C12Q1/6827 , C12Q1/6853 , C12Q1/686 , B01J2219/0059 , B01J2219/00722 , C40B40/06 , Y10S977/92
摘要： The present invention is directed to a process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof using a thermostable enzyme. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed.
摘要翻译： 本发明涉及使用热稳定性酶扩增核酸或其混合物中包含的任何靶核酸序列的方法。 该方法包括用摩尔过量的两种寡核苷酸引物处理核酸的分开的互补链，用热稳定酶扩增引物以形成互补引物延伸产物，其用作合成所需核酸序列的模板，并检测序列 放大 反应的步骤可以根据需要经常重复，并且涉及温度循环以进行杂交，促进酶的活性和所形成的杂交体的变性。
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公开(公告)号：US5804375A
公开(公告)日：1998-09-08
申请号：US428941
申请日：1995-04-25
CPC分类号： C12Q1/686 , C07H21/00 , C12Q1/6818 , C12Q1/6823 , C12Q1/6827 , C12Q1/703 , Y10S435/805 , Y10S435/81 , Y10S436/815
摘要： A process of detecting a target nucleic acid using labeled oligonucleotides uses the 5' to 3' nuclease activity of a nucleic acid polymerase to cleave annealed labeled oligonucleotide from hybridized duplexes and release labeled oligonucleotide fragments for detection. This process is easily incorporated into a PCR amplification assay.
摘要翻译： 使用标记的寡核苷酸检测靶核酸的过程使用核酸聚合酶的5'至3'核酸酶活性从杂交双链体切割退火标记的寡核苷酸，并释放标记的寡核苷酸片段进行检测。 该方法容易地并入PCR扩增测定中。
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公开(公告)号：US09102924B2
公开(公告)日：2015-08-11
申请号：US12425303
申请日：2009-04-16
申请人： Keith A. Bauer , Ellen Fiss , David H. Gelfand , Edward S. Smith , Shawn Suko , Olga Budker , Nancy Schoenbrunner , Susanne Stoffel , Thomas Myers
发明人： Keith A. Bauer , Ellen Fiss , David H. Gelfand , Edward S. Smith , Shawn Suko , Olga Budker , Nancy Schoenbrunner , Susanne Stoffel , Thomas Myers
IPC分类号： C12N9/12
CPC分类号： C12N9/1252 , C12N9/1276 , C12P19/34
摘要： Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. The mutant polymerases overcome the inhibitory effects by an intercalating dye. Therefore, the mutant polymerases are useful in a variety of disclosed methods in combination with an intercalating dye. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.
摘要翻译： 公开了相对于相应的未修饰聚合酶具有改进的延伸速率的突变型DNA聚合酶。 突变型聚合酶可用于各种公开的引物延伸方法。 突变型聚合酶克服了嵌入染料的抑制作用。 因此，突变型聚合酶可与各种公开的方法结合插入染料使用。 还公开了相关组合物，包括重组核酸，载体和宿主细胞，其可用于例如用于产生突变型DNA聚合酶。
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公开(公告)号：US20090280539A1
公开(公告)日：2009-11-12
申请号：US12425303
申请日：2009-04-16
申请人： Keith A. Bauer , Ellen Fiss , David H. Gelfand , Edward S. Smith , Shawn Suko , Olga Budker , Nancy Schoenbrunner , Susanne Stoffel
发明人： Keith A. Bauer , Ellen Fiss , David H. Gelfand , Edward S. Smith , Shawn Suko , Olga Budker , Nancy Schoenbrunner , Susanne Stoffel
CPC分类号： C12N9/1252 , C12N9/1276 , C12P19/34
摘要： Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. The mutant polymerases overcome the inhibitory effects by an intercalating dye. Therefore, the mutant polymerases are useful in a variety of disclosed methods in combination with an intercalating dye. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.
摘要翻译： 公开了相对于相应的未修饰聚合酶具有改进的延伸速率的突变型DNA聚合酶。 突变型聚合酶可用于各种公开的引物延伸方法。 突变型聚合酶克服了嵌入染料的抑制作用。 因此，突变型聚合酶可与各种公开的方法结合插入染料使用。 还公开了相关组合物，包括重组核酸，载体和宿主细胞，其可用于例如用于产生突变型DNA聚合酶。
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17.发明授权Recombinant expression vectors and purification methods for Thermus thermophilus DNA polymerase 失效Thermus thermophilus DNA聚合酶的重组表达载体和纯化方法
公开(公告)号：US5618711A
公开(公告)日：1997-04-08
申请号：US384490
申请日：1995-02-06
CPC分类号： C12N9/1252 , C12N9/1276
摘要： Recombinant DNA sequences encoding the DNA polymerase activity of Thermus thermophilus can be used to construct recombinant vectors and transformed host cells for production of the activity. T. thermophilus DNA polymerase is an .about.94 kDa protein especially useful in the DNA amplification procedure known as the polymerase chain reaction.
摘要翻译： 可以使用编码嗜热栖热菌的DNA聚合酶活性的重组DNA序列构建重组载体和转化的宿主细胞以产生该活性。 嗜热链球菌DNA聚合酶是DIFFERENCE 94 kDa蛋白质，特别适用于称为聚合酶链反应的DNA扩增过程。
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18.发明授权Mutated thermostable nucleic acid polymerase enzyme from thermotoga maritima 失效来自热带海洋假单胞菌的突变的热稳定性核酸聚合酶
公开(公告)号：US5420029A
公开(公告)日：1995-05-30
申请号：US971819
申请日：1993-02-03
IPC分类号： C12N1/21 , C12N9/12 , C12N15/00 , C12N15/09 , C12N15/54 , C12N15/70 , C12P19/34 , C12Q1/68 , C12Q1/70 , C12R1/01 , C12R1/19
CPC分类号： C12N9/1252 , C12N15/70 , C12N9/1276 , C12P19/34 , C12Q1/6876 , C12Q2521/101
摘要： A purified thermostable enzyme is derived from the eubacterium Thermotoga maritima. The enzyme has a molecular weight as determined by gel electrophoresis of about 97 kilodaltons and DNA polymerase I activity. The enzyme can be produced from native or recombinant host cells and can be used with primers and nucleoside triphosphates in a temperature-cycling chain reaction where at least one nucleic acid sequence is amplified in quantity from an existing sequence.
摘要翻译： PCT No.PCT / US91 / 05753 Sec。 371日期：1993年2月3日 102（e）日期1993年2月3日PCT 1991年8月13日PCT PCT。 出版物WO92 / 03556 日期1993年3月5日。纯化的热稳定酶源自海根杆菌。 该酶具有通过约97千道尔顿的凝胶电泳和DNA聚合酶I活性测定的分子量。 酶可以由天然或重组宿主细胞产生，并且可以在温度循环链反应中与引物和核苷三磷酸一起使用，其中至少一个核酸序列的量从现有序列中扩增。
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公开(公告)号：US4870013A
公开(公告)日：1989-09-26
申请号：US196789
申请日：1988-05-18
摘要： Expression vectors containing coding sequences under the control of SV40 early and RSV promoters are disclosed as useful in producing proteins in saccharomyces yeasts. Construction of such vectors, and their use in yeast transformations are described.
摘要翻译： 公开了含有SV40早期和RSV启动子控制下的编码序列的表达载体，用于在酵母菌中产生蛋白质。 描述了这些载体的构建及其在酵母转化中的应用。
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20.发明授权Purified thermostable nucleic acid polymerase enzyme from Thermotoga maritima 失效来自Thermotoga maritima的纯化的热稳定性核酸聚合酶
公开(公告)号：US5624833A
公开(公告)日：1997-04-29
申请号：US475231
申请日：1995-06-07
IPC分类号： C12N1/21 , C12N9/12 , C12N15/00 , C12N15/09 , C12N15/54 , C12N15/70 , C12P19/34 , C12Q1/68 , C12Q1/70 , C12R1/01 , C12R1/19
CPC分类号： C12N9/1252 , C12N15/70 , C12N9/1276 , C12P19/34 , C12Q1/6876 , C12Q2521/101
摘要： A purified thermostable enzyme is derived from the eubacterium Thermotoga maritima. The enzyme has a molecular weight as determined by gel electrophoresis of about 97 kilodaltons and DNA polymerase I activity. The enzyme can be produced from native or recombinant host cells and can be used with primers and nucleoside triphosphates in a temperature-cycling chain reaction where at least one nucleic acid sequence is amplified in quantity from an existing sequence.
摘要翻译： 纯化的热稳定性酶源自海绵杆菌（Thermotoga maritima）。 该酶具有通过约97千道尔顿的凝胶电泳和DNA聚合酶I活性测定的分子量。 酶可以由天然或重组宿主细胞产生，并且可以在温度循环链反应中与引物和核苷三磷酸一起使用，其中至少一个核酸序列的量从现有序列中扩增。
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