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1. 发明授权

US07973131B2 Extreme thermophile single-stranded DNA binding mutant protein, and nucleic acid isothermal amplification method of use thereof 有权
标题翻译：极性嗜热单链DNA结合突变蛋白，以及其使用的核酸等温扩增方法
公开(公告)号：US07973131B2
公开(公告)日：2011-07-05
申请号：US11495640
申请日：2006-07-31
申请人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
发明人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
IPC分类号： C07K1/00 , C07K14/00
CPC分类号： C07K14/195
摘要： The invention establishes a technology that allows non-specific amplification to be inhibited during nucleic acid amplification in an isothermal amplification reaction, such that the amplification efficiency is increased. The invention is a extreme thermophile single-stranded DNA binding mutant protein, having an amino acid sequence that expresses a function that can contribute to increasing an amplification efficiency of a template nucleic acid in an isothermal amplification reaction system that uses a strand displacement polymerase, and having in its amino acid sequence a mutation site where a mutation involving at least one of deletion, substitution, addition, and insertion of one or more amino acids in amino acid sequence of extreme thermophile single-stranded DNA binding protein has occurred, and a method of use thereof.
摘要翻译：本发明建立了在等温扩增反应的核酸扩增期间允许非特异性扩增被抑制的技术，使得扩增效率提高。本发明是一种极端嗜热单链DNA结合突变蛋白，其具有表达功能的氨基酸序列，该氨基酸序列可有助于增加使用链置换聚合酶的等温扩增反应系统中模板核酸的扩增效率，以及在其氨基酸序列中具有突变位点，其中发生了涉及极端嗜热性单链DNA结合蛋白的氨基酸序列中的一个或多个氨基酸的缺失，取代，添加和插入中的至少一个的突变，以及方法的使用。

2. 发明授权

US07713700B2 Method of amplifying nucleic acids, reagent kit for amplifying nucleic acids, method of detecting single nucleotide polymorphism, and reagent kit for detecting single nucleotide polymorphism 失效
标题翻译：核酸扩增方法，核酸扩增试剂盒，单核苷酸多态性检测方法，单核苷酸多态性检测试剂盒
公开(公告)号：US07713700B2
公开(公告)日：2010-05-11
申请号：US12062215
申请日：2008-04-03
申请人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa , Michio Oishi , Osamu Ohara
发明人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa , Michio Oishi , Osamu Ohara
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6848 , C12Q1/6858 , C12Q2521/507
摘要： An object of the present invention is to provide a nucleic acid amplification method for amplifying a desired nucleic acid while suppressing amplification of byproducts in a PCR reaction, a reagent kit used for nucleic acid amplification, a method of detecting single nucleotide polymorphism to detect single nucleotide polymorphism by utilizing that amplification of byproducts is suppressed in a PCR reaction, and a reagent kit used for detecting single nucleotide polymorphism. The method of amplifying nucleic acids by PCR is characterized by admixing in a reaction solution, a homologous recombinant protein which contains at least one of a RecA protein derived from Thermus thermophilus, and a modified RecA protein obtained by modification of the RecA protein and having a function similar to that of the RecA protein, and carrying out PCR.
摘要翻译：本发明的目的是提供用于扩增所需核酸同时抑制PCR反应中副产物扩增的核酸扩增方法，用于核酸扩增的试剂盒，检测单核苷酸多态性以检测单核苷酸的方法在PCR反应中抑制副产物扩增的多态性，以及用于检测单核苷酸多态性的试剂盒。通过PCR扩增核酸的方法的特征在于在反应溶液中混合含有至少一种源自Thermus thermophilus的RecA蛋白的同源重组蛋白，以及通过修饰RecA蛋白获得的经修饰的RecA蛋白，并且具有功能类似于RecA蛋白，并进行PCR。

3. 发明申请

US20060127978A1 Nucleic acid recombination method, host cell and expression vector 审中-公开
标题翻译：核酸重组方法，宿主细胞和表达载体
公开(公告)号：US20060127978A1
公开(公告)日：2006-06-15
申请号：US11071331
申请日：2005-03-04
申请人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
发明人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
IPC分类号： C12N15/09 , C12P21/06
CPC分类号： C12N15/902
摘要： This invention is to provide a nucleic acid recombination method which enables to carry out precisely homologous recombination of a desired target region, a host cell which enables to carry out precisely homologous recombination of a desired target region, and an expression vector which can be used for the nucleic acid recombination method and the host cell. The nucleic acid recombination method to carry out homologous recombination of a nucleic acid in a host cell employs a host cell which enables the expression and recombination of a RecA-like protein, and enables the expression of a RecX-like protein. And, the nucleic acid recombination method comprises a recombination step of a nucleic acid to carry out homologous recombination of the nucleic acid by the RecA-like protein expressed in the host cell, and a RecA inhibition step to inhibit the activity of the RecA-like protein by expressing the RecX-like protein.
摘要翻译：本发明提供一种核酸重组方法，其能够进行所需靶区域的精确同源重组，能够进行期望靶区域的精确同源重组的宿主细胞和可用于核酸重组方法和宿主细胞。用于在宿主细胞中进行核酸的同源重组的核酸重组方法使用能够表达和重组RecA样蛋白的宿主细胞，并且能够表达RecX样蛋白。并且，核酸重组方法包括核酸的重组步骤，以通过宿主细胞中表达的RecA样蛋白进行核酸的同源重组，以及抑制RecA样活性的RecA抑制步骤蛋白质通过表达RecX样蛋白。

4. 发明授权

US09303255B2 Electrode having enzyme crystals immobilized thereon, method for producing electrode having enzyme crystals immobilized thereon, and biological fuel cell and biosensor provided with electrode having enzyme crystals immobilized thereon 有权
标题翻译：在其上固定有酶晶体的电极，其上固定有酶晶体的电极的制备方法，以及生物燃料电池和具有固定有酶晶体的电极的生物传感器
公开(公告)号：US09303255B2
公开(公告)日：2016-04-05
申请号：US13818459
申请日：2011-08-24
申请人： Yasushi Shigemori , Yuichiro Nakaoki , Tsutomu Mikawa
发明人： Yasushi Shigemori , Yuichiro Nakaoki , Tsutomu Mikawa
IPC分类号： C12N11/06 , C12N9/04 , C12N9/02 , C12Q1/00 , H01M4/90 , H01M8/16 , C12N11/14 , C25B11/04 , H01M8/00
CPC分类号： C12N11/06 , C12N9/0006 , C12N9/0061 , C12N11/14 , C12Q1/001 , C12Y101/05002 , C25B11/0442 , H01M4/90 , H01M4/9008 , H01M8/00 , H01M8/16 , Y02E60/527 , Y02P70/56
摘要： An objective of the present invention is to establish a technique for making it possible to immobilize an enzyme on an electrically conductive base material in a uniformly, high density, and constantly aligned orientation, for the purpose of constructing an enzyme electrode having improved electrode performance. An electrode having enzyme crystals immobilized thereon, the electrode being provided with an electrically conductive base material that can be connected to an external circuit and enzyme crystals that serve as an electrode catalyst, wherein the enzyme crystals are immobilized on the electrically conducive base material; a method for producing an electrode having enzyme crystals immobilized thereon; and a biological fuel cell and a biosensor which are provided with an electrode having enzyme crystals immobilized thereon.
摘要翻译：本发明的目的是为了构建具有改善的电极性能的酶电极的目的，建立了可以以均匀，高密度和不断排列的取向将酶固定在导电性基材上的技术。具有固定在其上的酶结晶的电极，该电极设置有可连接到外部电路的导电基材和用作电极催化剂的酶晶体，其中所述酶晶体固定在所述导电基材上; 一种固定有酶晶体的电极的制造方法; 以及具有固定有酶晶体的电极的生物燃料电池和生物传感器。

5. 发明申请

US20070092896A1 Extreme thermophile single-stranded DNA binding mutant protein, and nucleic acid isothermal amplification method of use thereof 有权
标题翻译：极性嗜热单链DNA结合突变蛋白，以及其使用的核酸等温扩增方法
公开(公告)号：US20070092896A1
公开(公告)日：2007-04-26
申请号：US11495640
申请日：2006-07-31
申请人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
发明人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
IPC分类号： C12Q1/68 , C07H21/04 , C12P21/06 , C12N9/22 , C12N15/74 , C12N1/21
CPC分类号： C07K14/195
摘要： The invention establishes a technology that allows non-specific amplification to be inhibited during nucleic acid amplification in an isothermal amplification reaction, such that the amplification efficiency is increased. The invention is a extreme thermophile single-stranded DNA binding mutant protein, having an amino acid sequence that expresses a function that can contribute to increasing an amplification efficiency of a template nucleic acid in an isothermal amplification reaction system that uses a strand displacement polymerase, and having in its amino acid sequence a mutation site where a mutation involving at least one of deletion, substitution, addition, and insertion of one or more amino acids in amino acid sequence of extreme thermophile single-stranded DNA binding protein has occurred, and a method of use thereof.
摘要翻译：本发明建立了在等温扩增反应的核酸扩增期间允许非特异性扩增被抑制的技术，使得扩增效率提高。本发明是一种极端嗜热单链DNA结合突变蛋白，其具有表达功能的氨基酸序列，该氨基酸序列可有助于增加使用链置换聚合酶的等温扩增反应系统中模板核酸的扩增效率，以及在其氨基酸序列中具有突变位点，其中发生了涉及极端嗜热性单链DNA结合蛋白的氨基酸序列中的一个或多个氨基酸的缺失，取代，添加和插入中的至少一个的突变，以及方法的使用。

6. 发明申请

US20130224824A1 ELECTRODE HAVING ENZYME CRYSTALS IMMOBILIZED THEREON, METHOD FOR PRODUCING ELECTRODE HAVING ENZYME CRYSTALS IMMOBILIZED THEREON, AND BIOLOGICAL FUEL CELL AND BIOSENSOR PROVIDED WITH ELECTRODE HAVING ENZYME CRYSTALS IMMOBILIZED THEREON 有权
标题翻译：具有固定在其中的酶晶体的电极，用于生产其固定的酶的电极的方法和生物燃料电池和生物传感器，所述电极具有固定在其上的酶晶体的电极
公开(公告)号：US20130224824A1
公开(公告)日：2013-08-29
申请号：US13818459
申请日：2011-08-24
申请人： Yasushi Shigemori , Yuichiro Nakaoki , Tsutomu Mikawa
发明人： Yasushi Shigemori , Yuichiro Nakaoki , Tsutomu Mikawa
IPC分类号： C12N11/06 , H01M8/16 , H01M8/00 , C25B11/04
CPC分类号： C12N11/06 , C12N9/0006 , C12N9/0061 , C12N11/14 , C12Q1/001 , C12Y101/05002 , C25B11/0442 , H01M4/90 , H01M4/9008 , H01M8/00 , H01M8/16 , Y02E60/527 , Y02P70/56
摘要： An objective of the present invention is to establish a technique for making it possible to immobilize an enzyme on an electrically conductive base material in a uniformly, high density, and constantly aligned orientation, for the purpose of constructing an enzyme electrode having improved electrode performance. An electrode having enzyme crystals immobilized thereon, the electrode being provided with an electrically conductive base material that can be connected to an external circuit and enzyme crystals that serve as an electrode catalyst, wherein the enzyme crystals are immobilized on the electrically conducive base material; a method for producing an electrode having enzyme crystals immobilized thereon; and a biological fuel cell and a biosensor which are provided with an electrode having enzyme crystals immobilized thereon.
摘要翻译：本发明的目的是为了构建具有改善的电极性能的酶电极的目的，建立了可以以均匀，高密度和不断排列的取向将酶固定在导电性基材上的技术。具有固定在其上的酶结晶的电极，该电极设置有可连接到外部电路的导电基材和用作电极催化剂的酶晶体，其中所述酶晶体固定在所述导电基材上; 一种固定有酶晶体的电极的制造方法; 以及具有固定有酶晶体的电极的生物燃料电池和生物传感器。

7. 发明申请

US20080206775A1 Method of Amplifying Nucleic Acids, Reagent Kit for Amplifying Nucleic Acids, Method of Detecting Single Nucleotide Polymorphism, and Reagent Kit for Detecting Single Nucleotide Polymorphism 失效
标题翻译：扩增核酸的方法，扩增核酸的试剂盒，检测单核苷酸多态性的方法和用于检测单核苷酸多态性的试剂盒
公开(公告)号：US20080206775A1
公开(公告)日：2008-08-28
申请号：US12062215
申请日：2008-04-03
申请人： Yasushi SHIGEMORI , Takehiko SHIBATA , Tsutomu MIKAWA , Michio OISHI , Osamu OHARA
发明人： Yasushi SHIGEMORI , Takehiko SHIBATA , Tsutomu MIKAWA , Michio OISHI , Osamu OHARA
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6848 , C12Q1/6858 , C12Q2521/507
摘要： An object of the present invention is to provide a nucleic acid amplification method for amplifying a desired nucleic acid while suppressing amplification of byproducts in a PCR reaction, a reagent kit used for nucleic acid amplification, a method of detecting single nucleotide polymorphism to detect single nucleotide polymorphism by utilizing that amplification of byproducts is suppressed in a PCR reaction, and a reagent kit used for detecting single nucleotide polymorphism. The method of amplifying nucleic acids by PCR is characterized by admixing in a reaction solution, a homologous recombinant protein which contains at least one of a RecA protein derived from Thermus thermophilus, and a modified RecA protein obtained by modification of the RecA protein and having a function similar to that of the RecA protein, and carrying out PCR.
摘要翻译：本发明的目的是提供用于扩增所需核酸同时抑制PCR反应中副产物扩增的核酸扩增方法，用于核酸扩增的试剂盒，检测单核苷酸多态性以检测单核苷酸的方法在PCR反应中抑制副产物扩增的多态性，以及用于检测单核苷酸多态性的试剂盒。通过PCR扩增核酸的方法的特征在于在反应溶液中混合含有至少一种源自Thermus thermophilus的RecA蛋白的同源重组蛋白，以及通过修饰RecA蛋白获得的经修饰的RecA蛋白，并且具有功能类似于RecA蛋白，并进行PCR。

8. 发明申请

US20050277146A1 Method of amplifying template DNA molecule using strand-displacing DNA polymerase capable of carrying out isothermal amplification 审中-公开
标题翻译：使用能够进行等温扩增的链置换DNA聚合酶扩增模板DNA分子的方法
公开(公告)号：US20050277146A1
公开(公告)日：2005-12-15
申请号：US11138443
申请日：2005-05-27
申请人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
发明人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
IPC分类号： C12N15/09 , C12P19/34 , C12Q1/68
CPC分类号： C12Q1/6844 , C12Q2531/119 , C12Q2522/101
摘要： A method of amplifying a template DNA molecule in an isothermal reaction that can reduce background noise is provided. A method of amplifying a template DNA molecule using a strand-displacing DNA polymerase capable of carrying out isothermal amplification includes a step of conducting an amplification reaction with the addition of a single-strand DNA binding protein (SSB) from an extreme thermophile.
摘要翻译：提供了可以降低背景噪声的等温反应中扩增模板DNA分子的方法。使用能够进行等温扩增的链置换DNA聚合酶扩增模板DNA分子的方法包括从极端嗜热菌添加单链DNA结合蛋白（SSB）进行扩增反应的步骤。

9. 发明授权

US07902335B1 Heat-stable recA mutant protein and a nucleic acid amplification method using the heat-stable recA mutant protein 有权
标题翻译：热稳定recA突变蛋白和使用热稳定recA突变蛋白的核酸扩增方法
公开(公告)号：US07902335B1
公开(公告)日：2011-03-08
申请号：US11780284
申请日：2007-07-19
申请人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
发明人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa
IPC分类号： C07K14/00 , C07H21/02 , C12P21/04
CPC分类号： C12N9/00 , C12Q1/6844 , C12Q2521/507
摘要： A heat-stable RecA mutant protein is obtained by mutation involving either deletion or substitution of at least one amino acid in an amino acid sequence composing an acid region at C-terminal end of a wild type heat-stable RecA protein, and has an improved ability, compared to the wild type heat-stable RecA protein, for contributing to an increase in an amplification specificity of a template nucleic acid in a nucleic acid amplification reaction.
摘要翻译：通过涉及构成野生型热稳定性RecA蛋白C末端的酸性区域的氨基酸序列中的至少一个氨基酸的缺失或取代的突变获得热稳定性RecA突变蛋白，并且具有改良与野生型热稳定RecA蛋白相比，有助于增加模板核酸在核酸扩增反应中的扩增特异性。

10. 发明申请

US20050260631A1 Method of amplifying nucleic acids, reagent kit for amplifying nucleic acids, method of detecting single nucleotide polymorphism, and reagent kit for detecting single nucleotide polymorphism 审中-公开
标题翻译：核酸扩增方法，核酸扩增试剂盒，单核苷酸多态性检测方法，单核苷酸多态性检测试剂盒
公开(公告)号：US20050260631A1
公开(公告)日：2005-11-24
申请号：US11081949
申请日：2005-03-17
申请人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa , Michio Oishi , Osamu Ohara
发明人： Yasushi Shigemori , Takehiko Shibata , Tsutomu Mikawa , Michio Oishi , Osamu Ohara
IPC分类号： C12N15/09 , C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6848 , C12Q1/6858 , C12Q2521/507
摘要： An object of the present invention is to provide a nucleic acid amplification method for amplifying a desired nucleic acid while suppressing amplification of byproducts in a PCR reaction, a reagent kit used for nucleic acid amplification, a method of detecting single nucleotide polymorphism to detect single nucleotide polymorphism by utilizing that amplification of byproducts is suppressed in a PCR reaction, and a reagent kit used for detecting single nucleotide polymorphism. The method of amplifying nucleic acids by PCR is characterized by admixing in a reaction solution, a homologous recombinant protein which contains at least one of a RecA protein derived from Thermus thermophilus, and a modified RecA protein obtained by modification of the RecA protein and having a function similar to that of the RecA protein, and carrying out PCR.
摘要翻译：本发明的目的是提供用于扩增所需核酸同时抑制PCR反应中副产物扩增的核酸扩增方法，用于核酸扩增的试剂盒，检测单核苷酸多态性以检测单核苷酸的方法在PCR反应中抑制副产物扩增的多态性，以及用于检测单核苷酸多态性的试剂盒。通过PCR扩增核酸的方法的特征在于在反应溶液中混合含有至少一种源自Thermus thermophilus的RecA蛋白的同源重组蛋白，以及通过修饰RecA蛋白获得的经修饰的RecA蛋白，并且具有功能类似于RecA蛋白，并进行PCR。

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