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21.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：US5795762A
公开(公告)日：1998-08-18
申请号：US458819
申请日：1995-06-02
IPC分类号： C12N9/12
CPC分类号： C12N9/1252
摘要： The present invention relates to thermostable DNA polymerases which exhibit a different level of 5' to 3' exonuclease activity than their respective native polymerases. Particular conserved amino acid domains in thermostable DNA polymerases are mutated or deleted to alter the 5' to 3' exonuclease activity of the polymerases. The present invention also relates to means for isolating and producing such altered polymerases.
摘要翻译： 本发明涉及耐热性DNA聚合酶，其表现出与其各自的天然聚合酶不同的5'至3'核酸外切酶活性水平。 热稳定DNA聚合酶中特定的保守氨基酸结构域突变或缺失，以改变聚合酶的5'至3'核酸外切酶活性。 本发明还涉及用于分离和产生这种改变的聚合酶的方法。
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公开(公告)号：US5759828A
公开(公告)日：1998-06-02
申请号：US309512
申请日：1994-09-20
申请人： Rony Tal , David H. Gelfand , Roger D. Calhoon , Arie Ben-Bassat , Moshe Benziman , Hing Cheung Wong
发明人： Rony Tal , David H. Gelfand , Roger D. Calhoon , Arie Ben-Bassat , Moshe Benziman , Hing Cheung Wong
IPC分类号： C07K14/195 , C07K14/41 , C12N1/21 , C12N9/10 , C12N9/16 , C12N9/88 , C12N15/09 , C12N15/52 , C12P19/04 , C12P21/02 , C12R1/02 , C12N15/63 , C12N15/70 , C12N15/74 , C12P1/04
CPC分类号： C12N9/16 , C12N15/52 , C12N9/1051 , C12N9/88 , C12P19/04
摘要： The present invention provides the nucleotide sequences of Acetobacter operons, cdg operons encoding genes for the biosynthesis and degradation of cyclic diguanosine monophosphate (c-di-GMP). Specifically, the nucleotide sequences and deduced amino acid sequences of 3 phosphodiesterases isozymes, 3 diguanylate cyclase isozymes, and 2 polypeptides of unidentified function are provided. Also provided for are various strains of microorganisms, including Acetobacter cells genetically manipulated so as to produce elevated and/or reduced levels of one or more cdg operon encoded proteins.
摘要翻译： 本发明提供了醋酸杆菌操纵子的核苷酸序列，编码用于生物合成和环状二磷酸腺苷（c-di-GMP）的降解的基因的cdg操纵子。 具体地，提供了3种磷酸二酯酶同工酶，3个二环酸酯环化酶同功酶和2个未鉴定功能的多肽的核苷酸序列和推导的氨基酸序列。 还提供了各种微生物菌株，包括遗传操纵的醋杆菌细胞，以便产生升高和/或降低的一种或多种cdg操纵子编码的蛋白质的水平。
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24.发明授权
公开(公告)号：US5618703A
公开(公告)日：1997-04-08
申请号：US199509
申请日：1994-02-22
申请人： David H. Gelfand , Thomas W. Myers
发明人： David H. Gelfand , Thomas W. Myers
CPC分类号： C12Q1/703 , C12N15/70 , C12N9/1252 , C12N9/1276 , C12P19/34 , C12Q1/6827 , C12Q1/6844 , C12Q1/6848 , C12Q1/6853 , C12Q1/6858 , C12Q1/686 , C12Q1/6876
摘要： Methods are provided for distinguishing between RNA and DNA templates in an amplification reaction. In a preferred embodiment of the invention, the amplification reaction is a PCR and the reaction is catalyzed by a thermostable DNA polymerase or both reverse transcription and amplification of a target RNA. The invention particularly relates to selective amplification of RNA in the presence of homologous DNA, for example, HIV nucleic acids.
摘要翻译： 提供了用于区分扩增反应中的RNA和DNA模板的方法。 在本发明的一个优选实施方案中，扩增反应是PCR，并且反应由热稳定的DNA聚合酶或靶向RNA的逆转录和扩增两者催化。 本发明特别涉及在同源DNA例如HIV核酸存在下的RNA的选择性扩增。
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25.发明授权Methods for coupled high temperatures reverse transcription and polymerase chain reactions 失效用于偶联高温逆转录和聚合酶链反应的方法
公开(公告)号：US5561058A
公开(公告)日：1996-10-01
申请号：US449050
申请日：1995-05-24
CPC分类号： C12N9/1252 , C12N15/70 , C12N9/1276 , C12P19/34 , C12Q1/6848 , C12Q1/686
摘要： Methods are provided for the replication and amplification of RNA sequences by thermoactive DNA polymerases. In a preferred embodiment, high temperature reverse transcription is coupled to nucleic acid amplification in a one tube, one enzyme procedure using a thermostable DNA polymerase. Methods for eliminating carry over contamination of amplifications due to prior reverse transcription reactions are also provided. Reagents particularly suited for the methods of the present invention are provided.
摘要翻译： 提供了通过热活性DNA聚合酶复制和扩增RNA序列的方法。 在优选的实施方案中，将高温逆转录与一个管中的核酸扩增相结合，一种使用热稳定DNA聚合酶的酶程序。 还提供了用于消除由于先前的逆转录反应引起的扩增污染的方法。 提供特别适用于本发明方法的试剂。
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26.发明授权
公开(公告)号：US5374553A
公开(公告)日：1994-12-20
申请号：US567244
申请日：1990-08-13
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 , C12N1/20
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 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 temperaturecycling 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）。 该酶具有约97千道尔顿的分子量和DNA聚合酶I活性。 该酶可以由天然的或重组的宿主细胞产生，并且可以在温育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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27.发明授权Reverse transcription with thermostable DNA polymerase-high temperature reverse transcription 失效用热稳定DNA聚合酶 - 高温逆转录逆转录
公开(公告)号：US5310652A
公开(公告)日：1994-05-10
申请号：US82182
申请日：1993-06-24
申请人： David H. Gelfand , Thomas W. Myers
发明人： David H. Gelfand , Thomas W. Myers
CPC分类号： C12Q1/703 , C12N15/70 , C12N9/1252 , C12N9/1276 , C12P19/34 , C12Q1/6827 , C12Q1/6844 , C12Q1/6848 , C12Q1/6853 , C12Q1/6858 , C12Q1/686 , C12Q1/6876
摘要： Methods are provided for the replication and amplification of RNA sequences by thermoactive DNA polymerases. In a preferred embodiment, high temperature reverse transcription is coupled to nucleic acid amplification in a one tube, one enzyme procedure using a thermostable DNA polymerase. Methods for eliminating carry over contamination of amplifications due to prior reverse transcription reactions are also provided.
摘要翻译： 提供了通过热活性DNA聚合酶复制和扩增RNA序列的方法。 在优选的实施方案中，将高温逆转录与一个管中的核酸扩增相结合，一种使用热稳定DNA聚合酶的酶程序。 还提供了用于消除由于先前的逆转录反应引起的扩增污染的方法。
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28.发明授权Polypeptide expression using a portable temperature sensitive control cassette with a positive retroregulatory element 失效使用具有阳性逆转录调节元件的便携式温度敏感型控制盒的多肽表达
公开(公告)号：US4666848A
公开(公告)日：1987-05-19
申请号：US646709
申请日：1984-08-31
申请人： David H. Gelfand , Shing Chang , Hing C. Wong
发明人： David H. Gelfand , Shing Chang , Hing C. Wong
CPC分类号： C12N15/70 , C07K14/55 , C12N15/67 , C12N9/86 , Y10S435/849
摘要： The invention is a plasmid vector in which a first DNA sequence expressionable for a selected gene product is ligated to a positive retroregulatory element in a relationship thereto whereby expression of the selected gene product is enhanced. The first DNA sequence is furthermore operably linked to a second and third DNA sequence operably linked to one another which are respectively the P.sub.L promoter and a DNA sequence corresponding to an N gene ribosome binding site. Microorganisms transformed with the plasmid are also claimed.
摘要翻译： 本发明是一种质粒载体，其中将可选择的基因产物可表达的第一DNA序列与正向逆转录元件连接，从而提高所选择的基因产物的表达。 此外，第一DNA序列可操作地连接到彼此可操作地连接的第二和第三DNA序列，其分别是PL启动子和对应于N基因核糖体结合位点的DNA序列。 还要求用质粒转化的微生物。
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公开(公告)号：US07445900B2
公开(公告)日：2008-11-04
申请号：US11203461
申请日：2005-08-11
CPC分类号： C12Q1/686 , C07H21/00 , C12Q1/6818 , C12Q1/6823 , C12Q1/6827 , C12Q1/703 , Y10S435/805 , Y10S435/81 , Y10S436/815 , C12Q2521/101 , C12Q2565/1015 , C12Q2531/113 , C12Q2521/319 , C12Q2535/131 , C12Q2521/325 , C12Q2537/157 , C12Q2561/101 , C12Q2525/186 , C12Q2525/131
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：US20080171315A1
公开(公告)日：2008-07-17
申请号：US11203461
申请日：2005-08-11
IPC分类号： C12Q1/68
CPC分类号： C12Q1/686 , C07H21/00 , C12Q1/6818 , C12Q1/6823 , C12Q1/6827 , C12Q1/703 , Y10S435/805 , Y10S435/81 , Y10S436/815 , C12Q2521/101 , C12Q2565/1015 , C12Q2531/113 , C12Q2521/319 , C12Q2535/131 , C12Q2521/325 , C12Q2537/157 , C12Q2561/101 , C12Q2525/186 , C12Q2525/131
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.
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