公开(公告)号：US09890426B2

公开(公告)日：2018-02-13

申请号：US15439173

申请日：2017-02-22

申请人： Jingyue Ju ,  Zengmin Li ,  Sergey Kalachikov ,  Carl Fuller

发明人： Jingyue Ju ,  Zengmin Li ,  Sergey Kalachikov ,  Carl Fuller

IPC分类号： C12Q1/68 ,  C07K14/31 ,  C12N9/12 ,  C12N9/96

CPC分类号： C12Q1/6874 ,  A61K47/545 ,  A61K47/62 ,  A61K47/6889 ,  A61K47/69 ,  C07K14/31 ,  C12N9/1252 ,  C12N9/96 ,  C12Y207/07007

摘要： The present disclosure relates to compositions and methods based on a fast, efficient chemical reaction for conjugating a pore-forming protein, such as α-hemolysin, to a biomolecule, such as antibodies, receptors, and enzymes, such as DNA polymerase, and the use of such pore-forming protein conjugates in nanopore devices and methods.

公开(公告)号：US07393640B2

公开(公告)日：2008-07-01

申请号：US10772996

申请日：2004-02-05

申请人： Shiv Kumar ,  Mark McDougall ,  Anup Sood ,  John Nelson ,  Carl Fuller ,  John Macklin ,  Paul Mitsis

发明人： Shiv Kumar ,  Mark McDougall ,  Anup Sood ,  John Nelson ,  Carl Fuller ,  John Macklin ,  Paul Mitsis

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C07H19/10 ,  C07H19/20 ,  C07H21/00 ,  C12P19/34

摘要： The present invention describes methods of using terminal-phosphate-labeled nucleotides in the presence of a manganese salt to enhance their substrate properties towards various enzymes. Particularly described are methods of detecting a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrates for nucleic acid polymerases, in the presence of a manganese salt. Further provided are manganese complexes of terminal-phosphate-labeled nucleotides as well as terminal-phosphate-labeled nucleotides with new linkers with enhanced substrate properties.

摘要翻译： 本发明描述了在锰盐存在下使用末端磷酸酯标记的核苷酸以增强其对各种酶的底物性质的方法。 特别描述的是在锰盐存在的情况下，基于使用末端磷酸酯标记的核苷酸作为核酸聚合酶的底物，检测样品中的核酸的方法。 还提供了末端磷酸酯标记的核苷酸的锰络合物以及具有增强的底物性质的新接头的末端磷酸酯标记的核苷酸。

公开(公告)号：US07125671B2

公开(公告)日：2006-10-24

申请号：US10651362

申请日：2003-08-29

申请人： Anup Sood ,  Shiv Kumar ,  John Nelson ,  Carl Fuller ,  Anuradha Sekher

发明人： Anup Sood ,  Shiv Kumar ,  John Nelson ,  Carl Fuller ,  Anuradha Sekher

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6816 ,  C12Q1/6823 ,  C12Q1/6848 ,  C12Q1/686 ,  C12Q2525/101 ,  C12Q2521/525 ,  C12Q2521/101 ,  C12Q2565/301 ,  C12Q2561/113

摘要： The present invention relates generally to the use of terminal-phosphate-labeled nucleotides having three or more phosphates as substrates for nucleic acid polymerases and their use in DNA amplification. The labels employed are chemiluminescent, fluorescent, electrochemical and chromogenic moieties as well as mass tags and include those that are directly detectable, detectable after enzyme activation or feed into other processes to generate a different signal. The signal generated from the attached dyes may also be used to quantify the amount of amplification. Further provided are stabilizers that enhance the stability of terminal-phosphate labeled nucleoside polyphosphates in aqueous solutions and are useful for reducing non-enzymatic hydrolysis of these nucleotides, hence decrease background.

摘要翻译： 本发明一般涉及具有三种或更多种磷酸盐的末端磷酸酯标记的核苷酸作为核酸聚合酶的底物及其在DNA扩增中的应用。 使用的标记是化学发光，荧光，电化学和显色部分以及质量标签，并且包括直接可检测的标记，可在酶活化后进行或进入其它过程以产生不同信号。 从连接的染料产生的信号也可用于量化扩增量。 还提供了增强末端磷酸酯标记的核苷多磷酸酯在水溶液中的稳定性的稳定剂，并且可用于减少这些核苷酸的非酶水解，从而降低背景。

公开(公告)号：US20060147967A1

公开(公告)日：2006-07-06

申请号：US11315390

申请日：2005-12-22

申请人： Carl Fuller ,  Cuong Lam

发明人： Carl Fuller ,  Cuong Lam

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12P21/06 ,  C12N9/22 ,  C12N15/74

CPC分类号： C12Q1/6844 ,  C12N9/1252 ,  C12Q2531/119 ,  C12Q2521/101

摘要： A purified recombinant DNA polymerase having high processivity and strand-displacement activity. An isolated nucleic acid that encodes the bacteriophage DNA polymerase. A kit and method for amplifying DNA is also disclosed.

摘要翻译： 纯化的重组DNA聚合酶，其具有高的持续性和链置换活性。 编码噬菌体DNA聚合酶的分离核酸。 还公开了用于扩增DNA的试剂盒和方法。

公开(公告)号：US07052839B2

公开(公告)日：2006-05-30

申请号：US10113030

申请日：2002-04-01

申请人： John Nelson ,  Carl Fuller ,  Anup Sood ,  Shiv Kumar

发明人： John Nelson ,  Carl Fuller ,  Anup Sood ,  Shiv Kumar

IPC分类号： C12Q1/68

CPC分类号： C07H19/10 ,  C07H19/20 ,  C07H21/00 ,  C12Q1/6816 ,  C12Q1/6851

摘要： The present invention describes methods of detecting a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrates for nucleic acid polymerases. The methods provided by this invention utilize a nucleoside polyphosphate, dideoxynucleoside polyphosphate, or deoxynucleoside polyphosphate analogue which has a colorimetric dye, chemiluminescent, or fluorescent moiety, a mass tag or an electrochemical tag attached to the terminal-phosphate. When a nucleic acid polymerase uses this analogue as a substrate, an enzyme-activatable label would be present on the inorganic polyphosphate by-product of phosphoryl transfer. Cleavage of the polyphosphate product of phosphoryl transfer via phosphatase leads to a detectable change in the label attached thereon. When the polymerase assay is performed in the presence of a phosphatase, there is provided a convenient method for real-time monitoring of DNA or RNA synthesis and detection of a target nucleic acid.

公开(公告)号：US20180135121A1

公开(公告)日：2018-05-17

申请号：US15868593

申请日：2018-01-11

申请人： Jingyue Ju ,  Zengmin Li ,  Sergey Kalachikov ,  Carl Fuller

发明人： Jingyue Ju ,  Zengmin Li ,  Sergey Kalachikov ,  Carl Fuller

IPC分类号： C12Q1/6874 ,  C12N9/96 ,  C12N9/12 ,  C07K14/31

CPC分类号： C12Q1/6874 ,  A61K47/545 ,  A61K47/62 ,  A61K47/6889 ,  A61K47/69 ,  C07K14/31 ,  C07K19/00 ,  C12N9/1252 ,  C12N9/96 ,  C12Y207/07007

摘要： The present disclosure relates to compositions and methods based on a fast, efficient chemical reaction for conjugating a pore-forming protein, such as α-hemolysin, to a biomolecule, such as antibodies, receptors, and enzymes, such as DNA polymerase, and the use of such pore-forming conjugates in nanopore devices and methods.

公开(公告)号：US06875573B2

公开(公告)日：2005-04-05

申请号：US10190967

申请日：2002-07-08

申请人： Carl Fuller ,  Joseph Szasz

发明人： Carl Fuller ,  Joseph Szasz

IPC分类号： C12N9/12 ,  C12Q1/68 ,  C07H21/04

CPC分类号： C12N9/1252

摘要： Thermostable DNA polymerases both in native form and having single amino acid substitutions and optionally N-terminal deletions are disclosed. These polymerases exhibit a substantial improvement of DNA sequencing performance compared to Taq DNA polymerase. The instant DNA polymerases also possess improved salt tolerance.

摘要翻译： 公开了天然形式和具有单个氨基酸取代和任选的N-末端缺失的热稳定的DNA聚合酶。 与Taq DNA聚合酶相比，这些聚合酶显示出DNA测序性能的显着改善。 即时DNA聚合酶也具有改善的耐盐性。

公开(公告)号：US20170175183A1

公开(公告)日：2017-06-22

申请号：US15439173

申请日：2017-02-22

申请人： Jingyue Ju ,  Zengmin Li ,  Sergey Kalachikov ,  Carl Fuller

发明人： Jingyue Ju ,  Zengmin Li ,  Sergey Kalachikov ,  Carl Fuller

IPC分类号： C12Q1/68 ,  C12N9/12 ,  C12N9/96 ,  C07K14/31

CPC分类号： C12Q1/6874 ,  A61K47/545 ,  A61K47/62 ,  A61K47/6889 ,  A61K47/69 ,  C07K14/31 ,  C12N9/1252 ,  C12N9/96 ,  C12Y207/07007

摘要： The present disclosure relates to compositions and methods based on a fast, efficient chemical reaction for conjugating a pore-forming protein, such as α-hemolysin, to a biomolecule, such as antibodies, receptors, and enzymes, such as DNA polymerase, and the use of such pore-forming protein conjugates in nanopore devices and methods.

公开(公告)号：US07560254B2

公开(公告)日：2009-07-14

申请号：US10651558

申请日：2003-08-29

申请人： Anup Sood ,  Shiv Kumar ,  Carl Fuller ,  John Nelson ,  John Macklin

发明人： Anup Sood ,  Shiv Kumar ,  Carl Fuller ,  John Nelson ,  John Macklin

IPC分类号： C12P19/34 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04 ,  C07H21/00

CPC分类号： C12Q1/6853 ,  C12Q1/6858 ,  C12Q2525/125 ,  C12Q2521/525 ,  C12Q2521/319

摘要： A method of characterizing a nucleic acid sample is provided that includes the steps of: (a) conducting a DNA polymerase reaction that includes the reaction of a template, an allele specific primer, at least one terminal phosphate-labeled nucleotide, DNA polymerase, and optionally an enzyme having 3′→5′ exonuclease activity when the primer is non-hydrolyzable, which reaction results in the production of labeled polyphosphate; (b) permitting the labeled polyphosphate to react with a phosphatase to produce a detectable species; (c) detecting the detectable species; and (d) characterizing the nucleic acid sample based on such detection.

摘要翻译： 提供了表征核酸样品的方法，其包括以下步骤：（a）进行DNA聚合酶反应，其包括模板，等位基因特异性引物，至少一个末端磷酸酯标记的核苷酸，DNA聚合酶和 任选地当引物不可水解时具有3'→5'核酸外切酶活性的酶，该反应导致标记的多磷酸盐的产生; （b）允许标记的多磷酸酯与磷酸酶反应以产生可检测的物质; （c）检测可检测物种; 和（d）基于这种检测来表征核酸样品。

公开(公告)号：US20070292879A1

公开(公告)日：2007-12-20

申请号：US11781986

申请日：2007-07-24

申请人： ANUP SOOD ,  SHIV KUMAR ,  CARL FULLER ,  JOHN NELSON

发明人： ANUP SOOD ,  SHIV KUMAR ,  CARL FULLER ,  JOHN NELSON

IPC分类号： C12Q1/68

CPC分类号： C07H19/20 ,  C07H19/10 ,  C07H21/00 ,  C12Q1/6816 ,  C12Q1/6851 ,  C12Q1/6869 ,  C12Q2525/186

摘要： The present invention describes terminal phosphate blocked nucleoside polyphosphates that are stable at high temperature and their use in nucleic acid amplification and analysis. Current invention further describes charge modified terminal phosphate blocked nucleoside polyphosphates for improved incorporation and direct loading of nucleic acid sequencing reactions onto separating media.

摘要翻译： 本发明描述了在高温下稳定的磷酸封端的磷酸核苷多磷酸，并且其用于核酸扩增和分析。 本发明进一步描述了电荷修饰的末端磷酸酯封闭的核苷多磷酸盐，用于改进核酸测序反应在分离介质上的掺入和直接加载。