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1. 发明申请

US20060078928A1 Thermostable enzyme promoting the fidelity of thermostable DNA polymerases-for improvement of nucleic acid synthesis and amplification in vitro 有权
标题翻译：促进耐热DNA聚合酶保真度的热稳定酶 - 用于体外改进核酸合成和扩增
公开(公告)号：US20060078928A1
公开(公告)日：2006-04-13
申请号：US11241116
申请日：2005-09-29
申请人： Waltraud Ankenbauer , Frank Laue , Harald Sobek , Michael Greif
发明人： Waltraud Ankenbauer , Frank Laue , Harald Sobek , Michael Greif
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12N15/10 , C12N9/1252 , C12N9/22 , C12Q1/686 , C12Q2521/101
摘要： A purified thermostable enzyme is derived form the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3′-5′ exonuclease and cleaves to produce 5′-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Taq especially for nucleic acid amplification by the polymerase chain reaction (PCR).
摘要翻译：纯化的热稳定性酶源自嗜热古细菌大肠球菌（Archaeoglobus fulgidus）。酶可以是天然的或重组的，在PCR条件下是稳定的并且表现出双链特异性外切核酸酶活性。它是一个3'-5'核酸外切酶，并切割产生5'-单核苷酸。热稳定的核酸外切酶可用于许多重组DNA技术，与热稳定DNA聚合酶如Taq组合，特别是通过聚合酶链反应（PCR）进行核酸扩增。

2. 发明授权

US07410782B2 Thermostable enzyme promoting the fidelity of thermostable DNA polymerases—for improvement of nucleic acid synthesis and amplification in vitro 有权
标题翻译：促进耐热DNA聚合酶保真度的热稳定酶 - 用于体外改进核酸合成和扩增
公开(公告)号：US07410782B2
公开(公告)日：2008-08-12
申请号：US11241116
申请日：2005-09-29
申请人： Waltraud Ankenbauer , Frank Laue , Harald Sobek , Michael Greif
发明人： Waltraud Ankenbauer , Frank Laue , Harald Sobek , Michael Greif
IPC分类号： C12P19/34 , C12Q1/68 , C07K1/00 , C07H21/04
CPC分类号： C12N15/10 , C12N9/1252 , C12N9/22 , C12Q1/686 , C12Q2521/101
摘要： A purified thermostable enzyme is derived form the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3′-5′ exonuclease and cleaves to produce 5′-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Taq especially for nucleic acid amplification by the polymerase chain reaction (PCR).
摘要翻译：纯化的热稳定性酶源自嗜热古细菌大肠球菌（Archaeoglobus fulgidus）。酶可以是天然的或重组的，在PCR条件下是稳定的并且表现出双链特异性外切核酸酶活性。它是一个3'-5'核酸外切酶，并切割产生5'-单核苷酸。热稳定的核酸外切酶可用于许多重组DNA技术，与热稳定DNA聚合酶如Taq组合，特别是通过聚合酶链反应（PCR）进行核酸扩增。

3. 发明授权

US07030220B1 Thermostable enzyme promoting the fidelity of thermostable DNA polymerases-for improvement of nucleic acid synthesis and amplification in vitro 有权
标题翻译：促进耐热DNA聚合酶保真度的热稳定酶 - 用于体外改进核酸合成和扩增
公开(公告)号：US07030220B1
公开(公告)日：2006-04-18
申请号：US09856850
申请日：2000-09-27
申请人： Waltraud Ankenbauer , Frank Laue , Harald Sobek , Michael Greif
发明人： Waltraud Ankenbauer , Frank Laue , Harald Sobek , Michael Greif
IPC分类号： C07K1/00 , C12Q1/68 , C12P19/34 , C07H21/04
CPC分类号： C12N15/10 , C12N9/1252 , C12N9/22 , C12Q1/686 , C12Q2521/101
摘要： A purified thermostable enzyme is derived from the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3′–5′ exonuclease and cleaves to produce 5′-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Tag especially for nucleic acid amplification by the polymerase chain reaction (PCR).
摘要翻译：纯化的热稳定性酶源自嗜热古细菌大肠球菌（Archaeoglobus fulgidus）。酶可以是天然的或重组的，在PCR条件下是稳定的并且表现出双链特异性外切核酸酶活性。它是一个3'-5'核酸外切酶，并切割产生5'-单核苷酸。热稳定的核酸外切酶可用于许多重组DNA技术，与耐热性DNA聚合酶如Tag特异性用于通过聚合酶链反应（PCR）的核酸扩增。

4. 发明授权

US06607883B1 Polymerase chimeras 有权
标题翻译：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。

5. 发明授权

US07244602B2 Polymerase chimeras 有权
标题翻译：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。

6. 发明申请

US20120252071A1 T7 RNA POLYMERASE VARIANTS WITH CYSTEINE-SERINE SUBSTITUTIONS 审中-公开
标题翻译： 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。

7. 发明授权

US09193959B2 T7 RNA polymerase variants with enhanced thermostability 有权
标题翻译：具有增强热稳定性的T7 RNA聚合酶变体
公开(公告)号：US09193959B2
公开(公告)日：2015-11-24
申请号：US13069514
申请日：2011-03-23
申请人： Harald Sobek , Johann-Peter Thalhofer , Rainer Mueller , Manfred Schmidt , Michael Greif , Armin Ruf , Christian Rudolph
发明人： Harald Sobek , Johann-Peter Thalhofer , Rainer Mueller , Manfred Schmidt , Michael Greif , Armin Ruf , Christian Rudolph
IPC分类号： C12N9/12 , C12P19/34
CPC分类号： C12N9/1247 , C12P19/34 , C12Y207/07006
摘要： The present invention provides improved variants of T7 RNA polymerase by introducing novel mutations which lead to improved thermostability of the enzyme. According to the invention, amino acid substitutions at the positions Val426, Ser633, Val650, Thr654, Ala702, Val795, and combinations thereof are advantageous.
摘要翻译：本发明通过引入导致酶的热稳定性的新突变来提供T7RNA聚合酶的改进变体。根据本发明，位置Val426，Ser633，Val650，Thr654，Ala702，Val795及其组合的氨基酸取代是有利的。

8. 发明授权

US07378262B2 Reversibly modified thermostable enzymes for DNA synthesis and amplification in vitro 有权
标题翻译：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。

9. 发明申请

US20110256589A1 T7 RNA POLYMERASE VARIANTS WITH ENHANCED THERMOSTABILITY 有权
标题翻译：具有增强热稳定性的T7 RNA聚合酶变体
公开(公告)号：US20110256589A1
公开(公告)日：2011-10-20
申请号：US13069514
申请日：2011-03-23
申请人： Harald Sobek , Johann-Peter Thalhofer , Rainer Mueller , Manfred Schmidt , Michael Greif , Armin Ruf , Christian Rudolph
发明人： Harald Sobek , Johann-Peter Thalhofer , Rainer Mueller , Manfred Schmidt , Michael Greif , Armin Ruf , Christian Rudolph
IPC分类号： C12P19/34 , C12N5/10 , C12N15/63 , C12N9/10 , C07H21/00
CPC分类号： C12N9/1247 , C12P19/34 , C12Y207/07006
摘要： The present invention provides improved variants of T7 RNA polymerase by introducing novel mutations which lead to improved thermostability of the enzyme. According to the invention, amino acid substitutions at the positions Val426, Ser633, Val650, Thr654, Ala702, Val795, and combinations thereof are advantageous.
摘要翻译：本发明通过引入导致酶的热稳定性的新突变来提供T7RNA聚合酶的改进变体。根据本发明，位置Val426，Ser633，Val650，Thr654，Ala702，Val795及其组合的氨基酸取代是有利的。

10. 发明申请

US20110250598A1 DETERGENT FREE POLYMERASES 审中-公开
标题翻译：洗涤剂免费聚合物
公开(公告)号：US20110250598A1
公开(公告)日：2011-10-13
申请号：US13042628
申请日：2011-03-08
申请人： Ulrike Fischer , Michael Greif , Harald Sobek , Johann-Peter Thalhofer
发明人： Ulrike Fischer , Michael Greif , Harald Sobek , Johann-Peter Thalhofer
IPC分类号： C12Q1/68 , C12N9/12
CPC分类号： C12Q1/686 , C12N9/1252 , C12Q2527/137 , C12Q2521/101
摘要： The present invention relates to a formulation of a thermostable DNA polymerase which is completely free of detergents and its particular use in real time polymerase chain reaction (PCR). Such a formulation may be obtained if the selected purification method does not require the addition of a detergent at any purification step.
摘要翻译：本发明涉及完全不含洗涤剂的热稳定性DNA聚合酶的制剂及其在实时聚合酶链式反应（PCR）中的特别用途。如果所选择的纯化方法不需要在任何纯化步骤中添加洗涤剂，则可以获得这样的制剂。

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