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公开(公告)号：US20130034879A1
公开(公告)日：2013-02-07
申请号：US13564940
申请日：2012-08-02
CPC分类号： C12N9/1252 , C07K2319/21 , C12Y207/07007
摘要： A DNA polymerase mutant comprising a Taq DNA polymerase amino acid sequence with a mutation at one or more of the following selected amino acid positions: E189K, E230K, E507K, H28R, L30R, G38R, F73V, H75R, E76A, E76G, E76K, E90K, K206R, E315K, A348V, L351F, A439T, D452N, G504S, E507A, D551N, L552R, I553V, D578N, H676R, Q680R, D732G, E734G, E734K, F749V; wherein the polymerase mutant exhibits relative to wild-type DNA polymerase increased polymerase speed, increased affinity to DNA substrate and/or increased resistance to a DNA polymerase inhibitor; and wherein, when the mutation is E507K in combination with two or more further mutations or the mutation is Q680R in combination with four or more further mutations, at least one of the further mutations is at one of the selected amino acid positions; and when the mutation is I553V, this is not in combination with D551S.
摘要翻译： 一种DNA聚合酶突变体，其包含在一个或多个以下选择的氨基酸位置具有突变的Taq DNA聚合酶氨基酸序列：E189K，E230K，E507K，H28R，L30R，G38R，F73V，H75R，E76A，E76G，E76K，E90K ，K206R，E315K，A348V，L351F，A439T，D452N，G504S，E507A，D551N，L552R，I553V，D578N，H676R，Q680R，D732G，E734G，E734K，F749V; 其中所述聚合酶突变体相对于野生型DNA聚合酶显示出增加的聚合酶速度，对DNA底物的增加的亲和力和/或对DNA聚合酶抑制剂的增加的抗性; 并且其中当所述突变与两个或更多个其它突变组合时为E507K时，或者突变为Q680R与四个或更多个其它突变组合时，至少一个其它突变位于选定氨基酸位置之一; 当突变为I553V时，这不与D551S组合。
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公开(公告)号：US09493848B2
公开(公告)日：2016-11-15
申请号：US13564940
申请日：2012-08-02
申请人： Remigijus Skirgaila , Agne Tubelevi{hacek over (c)}iūté , Renata Rim{hacek over (s)}eliėnė , Sigitas Burinskas
发明人： Remigijus Skirgaila , Agne Tubelevi{hacek over (c)}iūté , Renata Rim{hacek over (s)}eliėnė , Sigitas Burinskas
IPC分类号： C12N9/12
CPC分类号： C12N9/1252 , C07K2319/21 , C12Y207/07007
摘要： The invention provides novel mutant thermostable DNA polymerases having desirable properties such as increased polymerase efficiency and speed, increased affinity to DNA substrates and/or increased resistance to DNA polymerase inhibitors. In certain embodiments, the invention provides methods of producing, amplifying and/or sequencing nucleic acid molecules (particularly cDNA molecules) using kits, compositions and/or reactions mixtures containing such novel DNA polymerases.
摘要翻译： 本发明提供具有所需性质的新型突变热稳定性DNA聚合酶，例如提高的聚合酶效率和速度，对DNA底物的增加的亲和力和/或增加的抗DNA聚合酶抑制剂。 在某些实施方案中，本发明提供使用包含这种新型DNA聚合酶的试剂盒，组合物和/或反应混合物来产生，扩增和/或测序核酸分子（特别是cDNA分子）的方法。
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公开(公告)号：US08835148B2
公开(公告)日：2014-09-16
申请号：US12899641
申请日：2010-10-07
CPC分类号： C12P19/34 , A61K38/162 , C12N9/1276 , C12N15/1075 , C12Y207/07049
摘要： The present invention provides a process for the production of nucleic acid encoding a target protein, which comprises: (a) providing an array of RNA or DNA molecules including one or more encoding the target protein; (b) generating a target protein from the array to form RNA-protein or DNA-protein complexes in which the RNA or DNA molecule is non-covalently or covalently bound to the complex; (c) separating the complexes into compartments wherein most or all of the compartments contain no more than one complex; (d) subjecting the complexes to reaction conditions which allow target protein activity; and (e) selecting nucleic acid encoding the target protein on the basis of the activity associated therewith, wherein when the complex is a DNA-protein complex in which the DNA is non-covalently bound, step b) is performed in the absence of separate compartments for each complex.
摘要翻译： 本发明提供了编码靶蛋白的核酸的制备方法，其包括：（a）提供包含一种或多种编码靶蛋白的RNA或DNA分子的阵列; （b）从阵列产生靶蛋白以形成其中RNA或DNA分子非共价或共价结合于复合物的RNA蛋白或DNA-蛋白复合物; （c）将配合物分离成隔室，其中大部分或全部隔室含有不超过一个络合物; （d）使复合物经受允许靶蛋白活性的反应条件; 基于与其相关的活性，选择编码靶蛋白的核酸，其中，当所述复合物是其中非共价结合DNA的DNA-蛋白复合物时，步骤b）在不存在分离的条件下进行 每个复合体的隔间。
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公开(公告)号：US09243233B2
公开(公告)日：2016-01-26
申请号：US13932365
申请日：2013-07-01
CPC分类号： C12Q1/6876 , C12N9/22 , C12Q1/683 , C12Q1/6883 , C12Q2600/154 , C12Q2600/158 , C12Q2521/331
摘要： A restriction endonuclease with a recognition sequence 5′-TCGA-3′. The restriction endonuclease is sensitive to the presence of a modified cytosine residue in the recognition sequence. Methods and kits using the restriction endonuclease with a recongition sequence 5′-TCGA-3′ are also disclosed.
摘要翻译： 具有识别序列5'-TCGA-3'的限制性内切核酸酶。 限制性内切酶对识别序列中修饰的胞嘧啶残基的存在敏感。 还公开了使用具有重组序列5'-TCGA-3'的限制性内切核酸酶的方法和试剂盒。
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公开(公告)号：US20110065606A1
公开(公告)日：2011-03-17
申请号：US12899641
申请日：2010-10-07
CPC分类号： C12P19/34 , A61K38/162 , C12N9/1276 , C12N15/1075 , C12Y207/07049
摘要： The present invention provides a process for the production of nucleic acid encoding a target protein, which comprises: (a) providing an array of RNA or DNA molecules including one or more encoding the target protein; (b) generating a target protein from the array to form RNA-protein or DNA-protein complexes in which the RNA or DNA molecule is non-covalently or covalently bound to the complex; (c) separating the complexes into compartments wherein most or all of the compartments contain no more than one complex; (d) subjecting the complexes to reaction conditions which allow target protein activity; and (e) selecting nucleic acid encoding the target protein on the basis of the activity associated therewith, wherein when the complex is a DNA-protein complex in which the DNA is non-covalently bound, step b) is performed in the absence of separate compartments for each complex.
摘要翻译： 本发明提供了编码靶蛋白的核酸的制备方法，其包括：（a）提供包含一种或多种编码靶蛋白的RNA或DNA分子的阵列; （b）从阵列产生靶蛋白以形成其中RNA或DNA分子非共价或共价结合于复合物的RNA蛋白或DNA-蛋白复合物; （c）将配合物分离成隔室，其中大部分或全部隔室含有不超过一个络合物; （d）使复合物经受允许靶蛋白活性的反应条件; 基于与其相关的活性，选择编码靶蛋白的核酸，其中当所述复合物是其中所述DNA非共价结合的DNA-蛋白复合物时，步骤b）在不存在分离的条件下进行 每个复合体的隔间。
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6.发明申请PHI29 DNA POLYMERASE MUTANTS HAVING INCREASED THERMOSTABILITY AND PROCESSIVITY 有权具有增加的热稳定性和加工性的PHI29 DNA聚合酶突变体公开(公告)号：US20140322759A1
公开(公告)日：2014-10-30
申请号：US14135860
申请日：2013-12-20
IPC分类号： C12N9/12
CPC分类号： C12N9/1252 , C12P19/34 , C12Y207/07007 , Y02P20/52
摘要： Mutants of bacteriophage phi29 DNA polymerase with increased protein stability and increased half-life, compared to wild type DNA polymerase. The disclosed mutants are more stable in reaction mixtures with or without DNA. The inventive phi29 DNA polymerase mutants generate more amplification product. The inventive phi29 DNA polymerase mutants amplify genomic DNA with less bias compared to wild type DNA polymerase. Selected mutations increase the affinity of polymerase for DNA template.
摘要翻译： 与野生型DNA聚合酶相比，噬菌体phi29 DNA聚合酶的突变体具有增加的蛋白质稳定性和增加的半衰期。 所公开的突变体在具有或不具有DNA的反应混合物中更稳定。 本发明的phi29 DNA聚合酶突变体产生更多的扩增产物。 与野生型DNA聚合酶相比，本发明的phi29DNA聚合酶突变体以较少的偏倚扩增基因组DNA。 所选择的突变增加了聚合酶对DNA模板的亲和力。
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公开(公告)号：US20140011196A1
公开(公告)日：2014-01-09
申请号：US13932365
申请日：2013-07-01
CPC分类号： C12Q1/6876 , C12N9/22 , C12Q1/683 , C12Q1/6883 , C12Q2600/154 , C12Q2600/158 , C12Q2521/331
摘要： A restriction endonuclease with a recognition sequence 5′-TCGA-3′. The restriction endonuclease is sensitive to the presence of a modified cytosine residue in the recognition sequence. Methods and kits using the restriction endonuclease with a recongition sequence 5′-TCGA-3′ are also disclosed.
摘要翻译： 具有识别序列5'-TCGA-3'的限制性内切核酸酶。 限制性内切酶对识别序列中修饰的胞嘧啶残基的存在敏感。 还公开了使用具有重组序列5'-TCGA-3'的限制性内切核酸酶的方法和试剂盒。
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公开(公告)号：US08580548B2
公开(公告)日：2013-11-12
申请号：US13408732
申请日：2012-02-29
IPC分类号： C12N9/12
CPC分类号： C12P19/34 , A61K38/162 , C12N9/1276 , C12N15/1075 , C12Y207/07049
摘要： The present invention provides a process for the production of nucleic acid encoding a target protein, which comprises: (a) providing an array of RNA or DNA molecules including one or more encoding the target protein; (b) generating a target protein from the array to form RNA-protein or DNA-protein complexes in which the RNA or DNA molecule is non-covalently or covalently bound to the complex; (c) separating the complexes into compartments wherein most or all of the compartments contain no more than one complex; (d) subjecting the complexes to reaction conditions which allow target protein activity; and (e) selecting nucleic acid encoding the target protein on the basis of the activity associated therewith, wherein when the complex is a DNA-protein complex in which the DNA is non-covalently bound, step b) is performed in the absence of separate compartments for each complex.
摘要翻译： 本发明提供了编码靶蛋白的核酸的制备方法，其包括：（a）提供包含一种或多种编码靶蛋白的RNA或DNA分子的阵列; （b）从阵列产生靶蛋白以形成其中RNA或DNA分子非共价或共价结合于复合物的RNA蛋白或DNA-蛋白复合物; （c）将配合物分离成隔室，其中大部分或全部隔室含有不超过一个络合物; （d）使复合物经受允许靶蛋白活性的反应条件; 基于与其相关的活性，选择编码靶蛋白的核酸，其中当所述复合物是其中所述DNA非共价结合的DNA-蛋白复合物时，步骤b）在不存在分离的条件下进行 每个复合体的隔间。
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