公开(公告)号：WO2011040971A3

公开(公告)日：2011-11-03

申请号：PCT/US2010002659

申请日：2010-09-30

Applicant: PACIFIC BIOSCIENCES CALIFORNIA ,  CLARK SONYA ,  BIBILLO AREK ,  PELUSO PAUL ,  CHRISTIANS FRED ,  HE MOLLY ,  PARK INSIL ,  LEE HAROLD ,  BJORNSON KEITH ,  JIA LEI ,  EMIG ROBIN

Inventor： CLARK SONYA ,  BIBILLO AREK ,  PELUSO PAUL ,  CHRISTIANS FRED ,  HE MOLLY ,  PARK INSIL ,  LEE HAROLD ,  BJORNSON KEITH ,  JIA LEI ,  EMIG ROBIN

IPC: C12N9/12 ,  C12N15/54 ,  C12Q1/68

CPC classification number: C12N9/1252 ,  C12Q1/6869 ,  C12Q2521/101

Abstract: Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions and/or heterologous or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include reduced reaction rates at one or more steps of the polymerase kinetic cycle, increased closed polymerase/DNA complex stability, enhanced metal ion coordination, reduced exonuclease activity, decreased branching fractions, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

公开(公告)号：WO2009145820A2

公开(公告)日：2009-12-03

申请号：PCT/US2009001974

申请日：2009-03-30

Applicant: PACIFIC BIOSCIENCES CALIFORNIA ,  CLARK SONYA ,  BIBILLO AREK ,  PELUSO PAUL ,  CHRISTIANS FRED ,  HE MOLLY ,  PARK INSIL ,  LEE HAROLD ,  BJORNSON KEITH ,  JIA LEI ,  EMIG ROBIN

Inventor： CLARK SONYA ,  BIBILLO AREK ,  PELUSO PAUL ,  CHRISTIANS FRED ,  HE MOLLY ,  PARK INSIL ,  LEE HAROLD ,  BJORNSON KEITH ,  JIA LEI ,  EMIG ROBIN

IPC: C12N9/12 ,  C12N15/10 ,  C12N15/54

CPC classification number: C12N9/1252 ,  C12Q1/6844 ,  C12Q1/6869 ,  C12Q2521/101

Abstract: Provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are less than the branching fractions of the polymerases from which they were derived, or branching fractions that are less than about 25% for a phosphate-labeled nucleotide analog. Also provided are compositions comprising modified recombinant polymerases that exhibit closed polymerase/DNA complexes with increased stability relative to the parental polymerases. Also provided are compositions comprising modified recombinant polymerases that exhibit decreased rate constants relative to the parental polymerases. Provided are methods for generating polymerases with the aforementioned phenotypes. Provided are methods of using such polymerases to make a DNA or to sequence a DNA template.

Abstract translation: 提供了包含修饰的重组聚合酶的组合物，其表现出小于其衍生自其的聚合酶的支化部分的支化部分，或对于磷酸酯标记的核苷酸类似物的支化部分小于约25％。 还提供了包含修饰的重组聚合酶的组合物，其表现出相对于亲本聚合酶具有增加的稳定性的闭合的聚合酶/ DNA复合物。 还提供了包含修饰的重组聚合酶的组合物，其相对于亲本聚合酶表现出降低的速率常数。 提供了产生具有上述表型的聚合酶的方法。 提供使用这种聚合酶制备DNA或顺序DNA模板的方法。

公开(公告)号：WO2011040971A2

公开(公告)日：2011-04-07

申请号：PCT/US2010002659

申请日：2010-09-30

Applicant: PACIFIC BIOSCIENCES CALIFORNIA ,  CLARK SONYA ,  BIBILLO AREK ,  PELUSO PAUL ,  CHRISTIANS FRED ,  HE MOLLY ,  PARK INSIL ,  LEE HAROLD ,  BJORNSON KEITH ,  JIA LEI ,  EMIG ROBIN

Inventor： CLARK SONYA ,  BIBILLO AREK ,  PELUSO PAUL ,  CHRISTIANS FRED ,  HE MOLLY ,  PARK INSIL ,  LEE HAROLD ,  BJORNSON KEITH ,  JIA LEI ,  EMIG ROBIN

IPC: C12N9/12 ,  C12N15/54 ,  C12Q1/68

CPC classification number: C12N9/1252 ,  C12Q1/6869 ,  C12Q2521/101

Abstract: Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions and/or heterologous or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include reduced reaction rates at one or more steps of the polymerase kinetic cycle, increased closed polymerase/DNA complex stability, enhanced metal ion coordination, reduced exonuclease activity, decreased branching fractions, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Abstract translation: 提供了包含重组DNA聚合酶的组合物，其包括氨基酸取代，插入，缺失和/或异源或外源特征，其在聚合酶上赋予改进的性质以用于增强的单分子测序。 这种性质可以包括聚合酶动力学循环的一个或多个步骤的降低的反应速率，增加的闭合聚合酶/ DNA复合物稳定性，增强的金属离子配位，减少的核酸外切酶活性，降低的支化部分等。 还提供了编码具有上述表型的聚合酶的核酸，以及使用这种聚合酶制备DNA或顺序DNA模板的方法。

公开(公告)号：EP2271751A4

公开(公告)日：2011-09-14

申请号：EP09755183

申请日：2009-03-30

Applicant: PACIFIC BIOSCIENCES CALIFORNIA

Inventor： CLARK SONYA ,  BIBILLO AREK ,  PELUSO PAUL ,  CHRISTIANS FRED ,  HE MOLLY ,  PARK INSIL ,  LEE HAROLD ,  BJORNSON KEITH ,  JIA LEI ,  EMIG ROBIN

IPC: C12Q1/68 ,  C12N9/12 ,  C12Q1/48

CPC classification number: C12N9/1252 ,  C12Q1/6844 ,  C12Q1/6869 ,  C12Q2521/101

公开(公告)号：EP1963536A4

公开(公告)日：2010-03-03

申请号：EP06848948

申请日：2006-12-21

Applicant: PACIFIC BIOSCIENCES CALIFORNIA

Inventor： HANZEL DAVID K ,  OTTO GEOFF ,  MURPHY DEVON ,  PELUSO PAUL ,  PHAM THANG ,  RANK DAVID R ,  MITSIS PAUL ,  CHRISTIANS FRED C ,  BIBILLO AREK ,  PARK INSIL ,  CLARK SONYA ,  LYLE JOHN

IPC: C12Q1/68

CPC classification number: C12N9/1252 ,  C07K2319/20 ,  C12P19/34 ,  C12Q1/6827 ,  C12Q1/6869 ,  C12Y207/07007 ,  Y02P20/52

Abstract: Compositions that include polymerases with features for improving entry of nucleotide analogues into active site regions and for coordinating with the nucleotide analogues in the active site region are provided. Methods of making the polymerases and of using the polymerases in sequencing and DNA replication and amplification as well as kinetic models of polymerase activity and computer-implemented methods of using the models are also provided.