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23 条记录 (耗时 0.031 秒)

    Polymerase chimeras
    2.
    发明授权
    Polymerase chimeras 有权
    聚合酶嵌合体

    公开(公告)号：US06607883B1

    公开(公告)日：2003-08-19

    申请号：US09623326

    申请日：2001-02-08

    申请人： Bruno Frey ,  Britta Villbrandt ,  Dietmar Schomburg ,  Harald Sobek ,  Waltraud Ankenbauer

    发明人： Bruno Frey ,  Britta Villbrandt ,  Dietmar Schomburg ,  Harald Sobek ,  Waltraud Ankenbauer

    IPC分类号： C12Q168

    CPC分类号： C12N9/1252 ,  C07K2319/00

    摘要： The invention concerns polymerase chimeras which are composed of amino acid fragments representing domains and which combine properties of naturally occurring polymerases that are advantageous with regard to a particular application. It has surprisingly turned out that the domains from the various enzymes are active in the chimeras and exhibit cooperative behavior. In addition the present invention concerns a process for the production of the chimeras according to the invention and the use of these chimeras for the synthesis of nucleic acids e.g. during a polymerase chain reaction. Moreover the present invention concerns a kit which contains the polymerase chimeras according to the invention.

    摘要翻译： 本发明涉及聚合酶嵌合体，其由表示结构域的氨基酸片段组成，并且其结合对于特定应用有利的天然存在的聚合酶的性质。 令人惊讶的是，各种酶的结构域在嵌合体中是活性的，并表现出合作行为。 此外，本发明涉及根据本发明的嵌合体的生产方法和这些嵌合体用于合成核酸的用途，例如， 在聚合酶链反应期间。 此外，本发明涉及包含根据本发明的聚合酶嵌合体的试剂盒。

    Polymerase chimeras
    3.
    发明授权
    Polymerase chimeras 有权
    聚合酶嵌合体

    公开(公告)号：US07244602B2

    公开(公告)日：2007-07-17

    申请号：US10456129

    申请日：2003-06-06

    申请人： Bruno Frey ,  Britta Villbrandt ,  Dietmar Schomburg ,  Harald Sobek ,  Waltraud Ankenbauer

    发明人： Bruno Frey ,  Britta Villbrandt ,  Dietmar Schomburg ,  Harald Sobek ,  Waltraud Ankenbauer

    IPC分类号： C12N9/12 ,  C12Q1/68

    CPC分类号： C12N9/1252 ,  C07K2319/00

    摘要： The invention concerns polymerase chimeras which are composed of amino acid fragments representing domains and which combine properties of naturally occurring polymerases that are advantageous with regard to a particular application. It has surprisingly turned out that the domains from the various enzymes are active in the chimeras and exhibit cooperative behavior. In addition the present invention concerns a process for the production of the chimeras according to the invention and the use of these chimeras for the synthesis of nucleic acids e.g. during a polymerase chain reaction. Moreover the present invention concerns a kit which contains the polymerase chimeras according to the invention.

    摘要翻译： 本发明涉及聚合酶嵌合体，其由表示结构域的氨基酸片段组成，并且其结合对于特定应用有利的天然存在的聚合酶的性质。 令人惊讶的是，各种酶的结构域在嵌合体中是活性的，并表现出合作行为。 此外，本发明涉及根据本发明的嵌合体的生产方法和这些嵌合体用于合成核酸的用途，例如， 在聚合酶链反应期间。 此外，本发明涉及包含根据本发明的聚合酶嵌合体的试剂盒。

    T7 RNA POLYMERASE VARIANTS WITH CYSTEINE-SERINE SUBSTITUTIONS
    4.
    发明申请
    T7 RNA POLYMERASE VARIANTS WITH CYSTEINE-SERINE SUBSTITUTIONS 审中-公开
    T7 RNA聚合酶与CYSTEINE-丝氨酸取代的变体

    公开(公告)号：US20120252071A1

    公开(公告)日：2012-10-04

    申请号：US13436110

    申请日：2012-03-30

    申请人： Michael Greif ,  Christian Rudolph ,  Manfred Schmidt ,  Harald Sobek ,  Johann-Peter Thalhofer

    发明人： Michael Greif ,  Christian Rudolph ,  Manfred Schmidt ,  Harald Sobek ,  Johann-Peter Thalhofer

    IPC分类号： C12P19/34 ,  C12N9/12

    CPC分类号： C12N9/1247 ,  C12Y207/07006

    摘要： The present disclosure provide novel variants of T7 RNA polymerase. Embodiments of T7 variants, according to the instant invention, include a Cysteine-Serine substitution on position 723 of the amino acid sequence of the T7 polypeptide. Embodiments of T7 variants according to the instant invention have a DNA-dependent RNA polymerase enzymatic activity and a reduced tendency to form intramolecular homodimers by way of oxidizing thiol groups. The amino acid substitutions within the T7 variants disclosed herein impact minimally, if at all, the RNA polymerase activity of the T7 polypeptide. Further, the mutations of the disclosed embodiments may optionally be combined with mutations which provide enhanced thermostability compared to the wild-type reference.

    摘要翻译： 本公开提供了T7RNA聚合酶的新变体。 根据本发明的T7变体的实施方案包括T7多肽的氨基酸序列的位置723上的半胱氨酸 - 丝氨酸取代。 根据本发明的T7变体的实施方案具有DNA依赖性RNA聚合酶酶活性，并且通过氧化硫醇基形成分子内同型二聚体的趋势降低。 本文公开的T7变体内的氨基酸取代最小程度地影响T7多肽的RNA聚合酶活性。 此外，所公开的实施方案的突变可以任选地与与野生型参考物相比提供增强的热稳定性的突变组合。

    Thermolabile uracil-DNA-glycosylas, process for its preparation and use for removing uracil from DNA
    5.
    发明授权
    Thermolabile uracil-DNA-glycosylas, process for its preparation and use for removing uracil from DNA 有权
    Thermolabile尿嘧啶-DNA-糖基，其制备方法和用于从DNA中去除尿嘧啶

    公开(公告)号：US06187575B1

    公开(公告)日：2001-02-13

    申请号：US09077312

    申请日：1998-08-06

    申请人： Harald Sobek ,  Manfred Schmidt ,  Bruno Frey ,  Klaus Kaluza

    发明人： Harald Sobek ,  Manfred Schmidt ,  Bruno Frey ,  Klaus Kaluza

    IPC分类号： C12N914

    CPC分类号： C12Q1/6848 ,  C12N9/2497 ,  C12Q2521/531

    摘要： Thermolabile enzyme with uracil-DNA-glycosylase activity which is in particular characterized by a high degree of purity, short half-lives and a content of contaminating foreign activities of less than 2%, a process for its isolation as well as the use thereof to remove the base uracil from DNA and in particular from PCR products containing uracil. The enzyme is obtainable from gram-positive microorganisms such as e.g. Arthrobacter or Micrococcus.

    摘要翻译： 具有尿嘧啶-DNA-糖基化酶活性的耐热不溶性酶，其特征在于其纯度高，半衰期短，外来污染物含量小于2％，其分离方法及其用途 从DNA中去除碱基尿嘧啶，特别是从含有尿嘧啶的PCR产物中除去。 该酶可从革兰氏阳性微生物获得，例如， 节杆菌属或微球菌属。

    Reversibly modified thermostable enzymes for DNA synthesis and amplification in vitro
    8.
    发明授权
    Reversibly modified thermostable enzymes for DNA synthesis and amplification in vitro 有权
    用于体外DNA合成和扩增的可逆修饰的热稳定酶

    公开(公告)号：US07378262B2

    公开(公告)日：2008-05-27

    申请号：US10317715

    申请日：2002-12-11

    申请人： Harald Sobek ,  Michael Greif

    发明人： Harald Sobek ,  Michael Greif

    IPC分类号： C12Q1/68 ,  C12N9/00 ,  C12N9/22

    CPC分类号： C12Q1/6848 ,  C12N9/1252 ,  C12N9/22 ,  C12N9/99 ,  C12Q1/686 ,  C12Q2549/101 ,  C12Q2521/319 ,  C12Q2521/101

    摘要： The invention relates to a composition comprising a first modified thermostable enzyme exhibiting 3′exonuclease activity but essentially no DNA polymerase activity and a second modified thermostable enzyme exhibiting DNA polymerase activity, whereas the fidelity of an amplification process is enhanced by the use of the composition in an amplification process in comparison to the use of the single second enzyme in an amplification process and, whereas said first and said second modified thermostable enzyme is reversibly modified by an inhibiting agent which results in essentially complete inactivation of enzyme activity, wherein incubation of said first and said second modified thermostable enzyme in an aqueous buffer at alkaline pH at a temperature less than 25° C. for 20 minutes results in no significant increase in the activity of said first and said second modified thermostable enzyme, wherein incubation at a temperature greater than 50° C. in an aqueous buffer at alkaline pH results in at least tow-fold increase in enzyme activity in less than 20 minutes which allow formation of primer extension products.

    摘要翻译： 本发明涉及一种组合物，其包含显示出3'核酸酶活性但基本上不含DNA聚合酶活性的第一个修饰的热稳定酶，以及显示DNA聚合酶活性的第二个修饰的热稳定酶，而通过使用组合物来增强扩增过程的保真度 扩增过程与在扩增过程中单次第二酶的使用相比较，而所述第一和所述第二修饰的热稳定酶由抑制剂可逆地修饰，所述抑制剂导致酶活性基本完全失活，其中所述第一 并且所述第二修饰的热稳定酶在碱性pH在低于25℃的温度下在水性缓冲液中20分钟导致所述第一和所述第二修饰的热稳定酶的活性没有显着增加，其中在大于 50℃，在碱性pH溶液中的水性缓冲液中 在少于20分钟的时间内至少可以提高酶活性的三倍增加，从而形成引物延伸产物。