公开(公告)号：US20090263813A1

公开(公告)日：2009-10-22

申请号：US12421188

申请日：2009-04-09

Applicant: DAVID H GELFAND ,  Ivo Glynne Gut ,  Keith A. Bauer ,  Florence Mauger

Inventor： DAVID H GELFAND ,  Ivo Glynne Gut ,  Keith A. Bauer ,  Florence Mauger

IPC: C12Q1/68

CPC classification number: C12Q1/6858 ,  C12Q1/686

Abstract: The present application provides polynucleotides comprising 5′-tails with sequence segments useful for the detection of target nucleic acid sequences, and methods for their use in detecting target nucleic acids. The polynucleotides are used to amplify a subsequence of a target nucleic acid in the presence of one or more ribonucleotides. The ribonucleotides are incorporated into amplification products at regular intervals complementary to the 5′-tail sequence segments. Cleavage of amplification products at the bond immediately 3′ to incorporated ribonucleotides produces detectably distinct fragments indicative of the presence or absence of a target nucleic acid.

Abstract translation: 本申请提供了包含具有用于检测靶核酸序列的序列区段的5'-尾的多核苷酸，以及它们用于检测靶核酸的方法。 多核苷酸用于在一个或多个核糖核苷酸存在下扩增靶核酸的亚序列。 核糖核苷酸以与5'-尾序列片段互补的规则间隔掺入扩增产物中。 立即将3'端的扩增产物切割成掺入的核糖核苷酸产生指示靶核酸存在或不存在的可检测的不同片段。

公开(公告)号：US20080293071A1

公开(公告)日：2008-11-27

申请号：US12158257

申请日：2006-12-20

Applicant: David H. Gelfand ,  Amar Gupta

Inventor： David H. Gelfand ,  Amar Gupta

IPC: C12Q1/68 ,  C12M1/00

CPC classification number: C12Q1/6869 ,  C12Q2525/186 ,  C12Q2521/525 ,  C12Q2521/319 ,  C12Q2521/301

Abstract: The invention provides a method of determining the nucleotide sequence of a target nucleic acid using a reversibly terminating nucleotide that is modified at the 2′ position.

Abstract translation: 本发明提供了使用在2'位置修饰的可逆终止的核苷酸来确定靶核酸的核苷酸序列的方法。

公开(公告)号：US06514736B1

公开(公告)日：2003-02-04

申请号：US09704357

申请日：2000-11-01

Applicant: Henry A. Erlich ,  Glenn Horn ,  Randall K. Saiki ,  Kary B. Mullis ,  David H. Gelfand

Inventor： Henry A. Erlich ,  Glenn Horn ,  Randall K. Saiki ,  Kary B. Mullis ,  David H. Gelfand

IPC: C12N912

CPC classification number: 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

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US6127155A

公开(公告)日：2000-10-03

申请号：US873897

申请日：1992-04-24

Applicant: David H. Gelfand ,  Susanne Stoffel ,  Randall K. Saiki

Inventor： David H. Gelfand ,  Susanne Stoffel ,  Randall K. Saiki

IPC: C12N9/12 ,  C12N15/70 ,  C12P19/34 ,  C12Q1/68 ,  C12N9/96 ,  C12N9/00 ,  C12P21/06

CPC classification number: C12P19/34 ,  C12N15/70 ,  C12N9/1252 ,  C12Q1/686 ,  Y10S435/814

Abstract: A purified thermostable nucleic acid polymerase is obtained that has unique characteristics. Preferably the nucleic acid polymerase is DNA polymerase isolated from a Thermus aquaticus species and has a molecular weight of about 86,000-95,000 daltons. The thermostable nucleic acid polymerase may be native or recombinant and may be used in a temperature-cycling chain reaction wherein at least one nucleic acid sequence is amplified in quantity from an existing sequence with the aid of selected primers and nucleotide triphosphates. The nucleic acid polymerase is preferably stored in a buffer containing non-ionic detergents that lends stability to the nucleic acid polymerase. A preferred buffer contains glycerol, polyoxyethylated sorbitan monolaurate, ethoxylated nonyl phenol and gelatin.

Abstract translation: 获得了具有独特特征的纯化的热稳定性核酸聚合酶。 优选地，所述核酸聚合酶是从栖热栖热菌属物种分离的DNA聚合酶，其分子量约为86,000-95,000道尔顿。 热稳定性核酸聚合酶可以是天然的或重组的，并且可以用于温度循环链式反应，其中借助于选定的引物和三磷酸核苷酸，从现存的序列中扩增至少一个核酸序列。 核酸聚合酶优选储存在含有使核酸聚合酶具有稳定性的非离子型清净剂的缓冲液中。 优选的缓冲液含有甘油，聚氧乙基化脱水山梨糖醇单月桂酸酯，乙氧基化壬基苯酚和明胶。

公开(公告)号：US6040166A

公开(公告)日：2000-03-21

申请号：US312729

申请日：1994-09-27

Applicant: Henry A. Erlich ,  Glenn Horn ,  Randall K. Saiki ,  Kary B. Mullis ,  David H. Gelfand

Inventor： Henry A. Erlich ,  Glenn Horn ,  Randall K. Saiki ,  Kary B. Mullis ,  David H. Gelfand

IPC: B01J19/00 ,  B01L7/00 ,  C07K14/74 ,  C07K14/805 ,  C12N9/12 ,  C12N15/10 ,  C12P19/34 ,  C12Q1/68 ,  C40B40/06

CPC classification number: 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

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US5795762A

公开(公告)日：1998-08-18

申请号：US458819

申请日：1995-06-02

Applicant: Richard D. Abramson ,  David H. Gelfand

Inventor： Richard D. Abramson ,  David H. Gelfand

IPC: C12N9/12

CPC classification number: C12N9/1252

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US5759828A

公开(公告)日：1998-06-02

申请号：US309512

申请日：1994-09-20

Applicant: Rony Tal ,  David H. Gelfand ,  Roger D. Calhoon ,  Arie Ben-Bassat ,  Moshe Benziman ,  Hing Cheung Wong

Inventor： Rony Tal ,  David H. Gelfand ,  Roger D. Calhoon ,  Arie Ben-Bassat ,  Moshe Benziman ,  Hing Cheung Wong

IPC: 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 classification number: C12N9/16 ,  C12N15/52 ,  C12N9/1051 ,  C12N9/88 ,  C12P19/04

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US5618703A

公开(公告)日：1997-04-08

申请号：US199509

申请日：1994-02-22

Applicant: David H. Gelfand ,  Thomas W. Myers

Inventor： David H. Gelfand ,  Thomas W. Myers

IPC: C12N9/12 ,  C12N15/70 ,  C12P19/34 ,  C12Q1/68 ,  C12Q1/70

CPC classification number: C12Q1/703 ,  C12N15/70 ,  C12N9/1252 ,  C12N9/1276 ,  C12P19/34 ,  C12Q1/6827 ,  C12Q1/6844 ,  C12Q1/6848 ,  C12Q1/6853 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q1/6876

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US5561058A

公开(公告)日：1996-10-01

申请号：US449050

申请日：1995-05-24

Applicant: David H. Gelfand ,  Thomas W. Myers ,  Christopher L. Sigua

Inventor： David H. Gelfand ,  Thomas W. Myers ,  Christopher L. Sigua

IPC: C12N9/12 ,  C12N15/70 ,  C12P19/34 ,  C12Q1/68 ,  C12Q1/70

CPC classification number: C12N9/1252 ,  C12N15/70 ,  C12N9/1276 ,  C12P19/34 ,  C12Q1/6848 ,  C12Q1/686

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US5374553A

公开(公告)日：1994-12-20

申请号：US567244

申请日：1990-08-13

Applicant: David H. Gelfand ,  Frances C. Lawyer

Inventor： David H. Gelfand ,  Frances C. Lawyer

IPC: 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 classification number: C12N9/1252 ,  C12N15/70 ,  C12N9/1276 ,  C12P19/34 ,  C12Q1/6876 ,  C12Q2521/101

Abstract: 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.

Abstract translation: 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。