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1. 发明授权

US08809048B2 Plasmid vector 有权
标题翻译：质粒载体
公开(公告)号：US08809048B2
公开(公告)日：2014-08-19
申请号：US13637945
申请日：2011-03-18
申请人： Yukari Ohta , Yuji Hatada , Kozue Mori , Nobuyuki Nakamura
发明人： Yukari Ohta , Yuji Hatada , Kozue Mori , Nobuyuki Nakamura
IPC分类号： C12N15/00 , C12N1/00 , C12N1/20 , C12P21/04 , C12P21/06
CPC分类号： C12N15/74
摘要： The object is to provide a technique for modifying a bacterium belonging to the genus Kocuria through genetic engineering for industrially effectively utilizing the bacterium belonging to the genus Kocuria. The above object can be achieved by providing a cyclic plasmid, which has a replication region comprising the base sequence of a DNA-binding protein-like protein gene, the base sequence of a replicase-like protein gene, and the base sequence represented by SEQ ID NO:45, and is autonomously replicable in bacteria. As an example of the aforesaid plasmid, a plasmid containing a base sequence represented by SEQ ID NO:3 or 4 which originates in Kocuria sp. MBE131 strain (FERM P-21885) can be cited.
摘要翻译：本发明的目的是提供一种用于通过遗传工程修饰属于Kocuria属的细菌的技术，用于工业上有效利用属于Kocuria属的细菌。上述目的可以通过提供一种环状质粒来实现，该质粒具有包含DNA结合蛋白样蛋白质基因的碱基序列，复制酶样蛋白质基因的碱基序列和SEQ ID ID NO：45，并且在细菌中是可自主复制的。作为上述质粒的实例，含有由SEQ ID NO：3或4表示的碱基序列的质粒，其起源于Kocuria sp。可引用MBE131菌株（FERM P-21885）。

2. 发明申请

US20130189760A1 NOVEL EXTRACELLULARLY SECRETED NUCLEASE 有权
标题翻译：新颖的细胞分泌核酸
公开(公告)号：US20130189760A1
公开(公告)日：2013-07-25
申请号：US13823990
申请日：2011-09-15
申请人： Kozue Mori , Yukari Ohta , Yuji Hatada , Nobuyuki Nakamura , Masayuki Miyazaki
发明人： Kozue Mori , Yukari Ohta , Yuji Hatada , Nobuyuki Nakamura , Masayuki Miyazaki
IPC分类号： C12N9/22 , C12N1/08 , C12R1/465
CPC分类号： C12N9/22 , C12N1/08 , C12Q1/44 , C12R1/465
摘要： The purpose of the present invention is to provide a nuclease that secretes natural nonpathogenic microorganisms extracellularly, has higher specific activity than conventional nucleases, and is useful in nucleolytic degradation on an industrial scale. This purpose is achieved with an extracellularly secreted nuclease derived from Streptomyces bacteria, the nuclease having specific activity equal to or greater than the specific activity of Benzonase® when supplied to double-stranded DNA for 30 minutes at 37° C. in 20 mM Tris/HCl (pH 8.5) containing 1 mM MgCl2 and 1 mM CaCl2 after purification, using double-stranded DNA, single-stranded DNA, and RNA as substrates.
摘要翻译：本发明的目的是提供一种在细胞外分泌天然非致病微生物的核酸酶，比常规核酸酶具有更高的比活性，并且可用于工业规模的核酸分解降解。该目的通过来自链霉菌细菌的细胞外分泌的核酸酶实现，所述核酸酶在37℃下在20mM Tris / HCl中提供至双链DNA 30分钟时具有等于或大于Benzonase的比活性的比活性，纯化后含有1mM MgCl 2和1mM CaCl 2的HCl（pH8.5），使用双链DNA，单链DNA和RNA作为底物。

3. 发明申请

US20130084621A1 PLASMID VECTOR 有权
公开(公告)号：US20130084621A1
公开(公告)日：2013-04-04
申请号：US13637945
申请日：2011-03-18
申请人： Yukari Ohta , Yuji Hatada , Kozue Mori , Nobuyuki Nakamura
发明人： Yukari Ohta , Yuji Hatada , Kozue Mori , Nobuyuki Nakamura
IPC分类号： C12N15/74
CPC分类号： C12N15/74
摘要： The object is to provide a technique for modifying a bacterium belonging to the genus Kocuria through genetic engineering for industrially effectively utilizing the bacterium belonging to the genus Kocuria. The above object can be achieved by providing a cyclic plasmid, which has a replication region comprising the base sequence of a DNA-binding protein-like protein gene, the base sequence of a replicase-like protein gene, and the base sequence represented by SEQ ID NO:45, and is autonomously replicable in bacteria. As an example of the aforesaid plasmid, a plasmid containing a base sequence represented by SEQ ID NO:3 or 4 which originates in Kocuria sp. MBE131 strain (FERM P-21885) can be cited.
摘要翻译：本发明的目的是提供一种用于通过遗传工程修饰属于Kocuria属的细菌的技术，用于工业上有效利用属于Kocuria属的细菌。上述目的可以通过提供一种环状质粒来实现，该质粒具有包含DNA结合蛋白样蛋白质基因的碱基序列，复制酶样蛋白质基因的碱基序列和SEQ ID ID NO：45，并且在细菌中是可自主复制的。作为上述质粒的实例，含有由SEQ ID NO：3或4表示的碱基序列的质粒，其起源于Kocuria sp。可引用MBE131菌株（FERM P-21885）。

4. 发明授权

US09217139B2 Extracellularly secreted nuclease 有权
标题翻译：细胞外分泌的核酸酶
公开(公告)号：US09217139B2
公开(公告)日：2015-12-22
申请号：US13823990
申请日：2011-09-15
申请人： Kozue Mori , Yukari Ohta , Yuji Hatada , Nobuyuki Nakamura , Masayuki Miyazaki
发明人： Kozue Mori , Yukari Ohta , Yuji Hatada , Nobuyuki Nakamura , Masayuki Miyazaki
IPC分类号： C12N9/22 , C12R1/465 , C12Q1/44 , C12N1/08
CPC分类号： C12N9/22 , C12N1/08 , C12Q1/44 , C12R1/465
摘要： The purpose of the present invention is to provide a nuclease that secretes natural nonpathogenic microorganisms extracellularly, has higher specific activity than conventional nucleases, and is useful in nucleolytic degradation on an industrial scale. This purpose is achieved with an extracellularly secreted nuclease derived from Streptomyces bacteria, the nuclease having specific activity equal to or greater than the specific activity of Benzonase® when supplied to double-stranded DNA for 30 minutes at 37° C. in 20 mM Tris/HCl (pH 8.5) containing 1 mM MgCl2 and 1 mM CaCl2 after purification, using double-stranded DNA, single-stranded DNA, and RNA as substrates.
摘要翻译：本发明的目的是提供一种在细胞外分泌天然非致病微生物的核酸酶，比常规核酸酶具有更高的比活性，并且可用于工业规模的核酸分解降解。该目的通过来自链霉菌细菌的细胞外分泌的核酸酶实现，所述核酸酶在37℃下在20mM Tris / HCl中提供至双链DNA 30分钟时具有等于或大于Benzonase的比活性的比活性，纯化后含有1mM MgCl 2和1mM CaCl 2的HCl（pH8.5），使用双链DNA，单链DNA和RNA作为底物。

5. 发明申请

US20110008832A1 METHODS FOR STABLY RETAINING FOREIGN GENES IN CELLS 审中-公开
标题翻译：在细胞中稳定保留外源基因的方法
公开(公告)号：US20110008832A1
公开(公告)日：2011-01-13
申请号：US12452645
申请日：2008-07-09
申请人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
发明人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
IPC分类号： C12P21/00 , C12N15/63 , C12N1/21 , C12N1/16 , C12N5/10
CPC分类号： C12N15/64 , C12N9/93 , C12N15/821
摘要： A method for preparing a protein or peptide encoded by a foreign gene by expressing a foreign gene, which comprises the steps of: preparing a recombinant vector comprising in an expressible state a gene encoding an aminoacyl-tRNA synthetase, in which a desired foreign gene has been inserted in an expressible state; preparing a mutant host cell or the like in which a chromosomal gene encoding an aminoacyl-tRNA synthetase has been knocked out; transforming the mutant host cell with the recombinant vector to obtain a transformant; and culturing the transformant to prepare the protein or peptide encoded by the foreign gene. It becomes possible to provide a novel means for permitting the retention of a recombinant DNA cloning vector without employing an antibiotic and without limiting the composition of the medium; and a method for preparing a protein or peptide encoded by a foreign gene by using the means.
摘要翻译：一种通过表达外源基因来制备由外来基因编码的蛋白质或肽的方法，该方法包括以下步骤：制备包含可表达状态的编码氨基-tRNA合成酶的基因的重组载体，其中期望的外源基因具有被插入可表达的状态; 制备其中编码氨基-tRNA合成酶的染色体基因已经被敲除的突变宿主细胞等; 用重组载体转化突变宿主细胞以获得转化体; 并培养转化体以制备由外源基因编码的蛋白质或肽。可以提供一种用于在不使用抗生素的情况下保留重组DNA克隆载体并且不限制培养基的组成的新方法; 以及通过使用该方法制备由外源基因编码的蛋白质或肽的方法。

6. 发明申请

US20110014677A1 NOVEL DNA FRAGMENT, RECOMBINANT VECTOR COMPRISING SAME, TRANSFORMANT TRANSFORMED THEREWITH, AND USE THEREOF 失效
标题翻译：新型DNA片段，包含其的重组载体，其变形变体及其用途
公开(公告)号：US20110014677A1
公开(公告)日：2011-01-20
申请号：US12452648
申请日：2008-07-09
申请人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
发明人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
IPC分类号： C12N9/02 , C12N9/10 , C12N9/14 , C12N9/88 , C12N9/90 , C12N1/21 , C12N15/63 , C07H21/04
CPC分类号： C12N15/63 , C07K2319/02 , C12N9/2437 , C12N9/2468 , C12N15/70 , C12N15/75 , C12P21/02 , C12Y302/01004 , C12Y302/01081
摘要： A DNA fragment containing a gene which encodes a specific gene regulatory region alone or the gene regulatory region together with a signal peptide; a recombinant vector containing the DNA fragment; a transformant containing the recombinant vector; and a method of producing a recombinant protein by using the transformant. According to the invention, it is possible to produce a protein in a large amount at a high efficiency regardless of the kind of the recombinant protein.
摘要翻译：含有单独编码特定基因调控区域的基因或与信号肽一起的基因调控区域的DNA片段; 含有该DNA片段的重组载体; 含有重组载体的转化体; 以及使用该转化体生产重组蛋白的方法。根据本发明，无论重组蛋白质的种类如何，都可以高效率地生产蛋白质。

7. 发明授权

US08354248B2 Promoter-encoding DNA fragment, recombinant vector, recombiant transformant, and uses thereof 失效
标题翻译：启动子编码DNA片段，重组载体，重组转化体及其用途
公开(公告)号：US08354248B2
公开(公告)日：2013-01-15
申请号：US12452648
申请日：2008-07-09
申请人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
发明人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
IPC分类号： C12P21/06 , C12N9/00 , C12N9/02 , C12N9/10 , C12N9/88 , C12N1/20 , C12N15/00 , C07H21/04
CPC分类号： C12N15/63 , C07K2319/02 , C12N9/2437 , C12N9/2468 , C12N15/70 , C12N15/75 , C12P21/02 , C12Y302/01004 , C12Y302/01081
摘要： A DNA fragment containing a gene which encodes a specific gene regulatory region alone or the gene regulatory region together with a signal peptide; a recombinant vector containing the DNA fragment; a transformant containing the recombinant vector; and a method of producing a recombinant protein by using the transformant. According to the invention, it is possible to produce a protein in a large amount at a high efficiency regardless of the kind of the recombinant protein.
摘要翻译：含有单独编码特定基因调控区域的基因或与信号肽一起的基因调控区域的DNA片段; 含有该DNA片段的重组载体; 含有重组载体的转化体; 以及使用该转化体生产重组蛋白的方法。根据本发明，无论重组蛋白质的种类如何，都可以高效率地生产蛋白质。

8. 发明授权

US08790907B2 Method for detachment and preparation of living microorganisms 有权
标题翻译：活细菌分离和制备方法
公开(公告)号：US08790907B2
公开(公告)日：2014-07-29
申请号：US13359797
申请日：2012-01-27
申请人： Sumihiro Koyama , Tadashi Maruyama , Chiaki Kato , Yuichi Nogi , Yuji Hatada , Yukari Ohta , Masaaki Konishi , Taishi Tsubouchi
发明人： Sumihiro Koyama , Tadashi Maruyama , Chiaki Kato , Yuichi Nogi , Yuji Hatada , Yukari Ohta , Masaaki Konishi , Taishi Tsubouchi
IPC分类号： C12N1/00 , C12N1/04 , C12N11/14
CPC分类号： C12N13/00 , C12N11/14
摘要： A method for immobilizing living microorganisms includes a step (1) of disposing a solution containing microorganisms as an electrolyte on the surface of a substrate at least one portion of which is an electrode, and applying a constant potential to the electrode to cause at least a portion of the microorganisms to attach to the surface of the substrate. The constant potential in step (1) is greater than −0.5 V but not greater than −0.2 V (vs Ag/AgCl) or greater than +0.2 V but not greater than +0.4 V (vs Ag/AgCl). The electrolyte in step (1) does not contain a source of nutrition for the microorganisms.
摘要翻译：固定活体微生物的方法包括将含有微生物作为电解质的溶液设置在基材的表面上的步骤（1），其中至少一部分是电极，并向电极施加恒定电位，使至少部分微生物附着到基底表面。步骤（1）中的恒定电位大于-0.5V但不大于-0.2V（相对于Ag / AgCl）或大于+ 0.2V但不大于+0.4V（相对于Ag / AgCl）。步骤（1）中的电解质不含微生物的营养来源。

9. 发明申请

US20130196404A1 METHOD FOR DETACHMENT AND PREPARATION OF LIVING MICROORGANISMS 有权
标题翻译：生活微生物的分离和制备方法
公开(公告)号：US20130196404A1
公开(公告)日：2013-08-01
申请号：US13359797
申请日：2012-01-27
申请人： Sumihiro Koyama , Tadashi Maruyama , Chiaki Kato , Yuichi Nogi , Yuji Hatada , Yukari Ohta , Masaaki Konishi , Taishi Tsubouchi
发明人： Sumihiro Koyama , Tadashi Maruyama , Chiaki Kato , Yuichi Nogi , Yuji Hatada , Yukari Ohta , Masaaki Konishi , Taishi Tsubouchi
IPC分类号： C12N13/00
CPC分类号： C12N13/00 , C12N11/14
摘要： A method for immobilizing living microorganisms includes a step (1) of disposing a solution containing microorganisms as an electrolyte on the surface of a substrate at least one portion of which is an electrode, and applying a constant potential to the electrode to cause at least a portion of the microorganisms to attach to the surface of the substrate. The constant potential in step (1) is greater than −0.5 V but not greater than −0.2 V (vs Ag/AgCl) or greater than +0.2 V but not greater than +0.4 V (vs Ag/AgCl). The electrolyte in step (1) does not contain a source of nutrition for the microorganisms.
摘要翻译：固定活体微生物的方法包括将含有微生物作为电解质的溶液设置在基材的表面上的步骤（1），其中至少一部分是电极，并向电极施加恒定电位，使至少部分微生物附着到基底表面。步骤（1）中的恒定电位大于-0.5V但不大于-0.2V（相对于Ag / AgCl）或大于+ 0.2V但不大于+0.4V（相对于Ag / AgCl）。步骤（1）中的电解质不含微生物的营养来源。

10. 发明授权

US06689874B2 Gene for enzyme having both alkaline pullulanase and alkaline alpha-amylase activities 失效
标题翻译：具有碱性支链淀粉酶和碱性α-淀粉酶活性的酶的基因
公开(公告)号：US06689874B2
公开(公告)日：2004-02-10
申请号：US10014436
申请日：2001-12-14
申请人： Yuji Hatada , Kazuaki Igarashi , Katsuya Ozaki , Katsutoshi Ara , Shuji Kawai , Susumu Ito
发明人： Yuji Hatada , Kazuaki Igarashi , Katsuya Ozaki , Katsutoshi Ara , Shuji Kawai , Susumu Ito
IPC分类号： C07H2104
CPC分类号： C12N9/2457 , C12N9/2417 , C12N9/2451 , C12Y302/01001 , C12Y302/01041
摘要： The present invention provides: a nucleotide sequence encoding alkaline pullulanase and exhibiting both alkaline &agr;-amylase and alkaline pullulanase activity; a nucleotide sequence encoding alkaline &agr;-amylase possessing an amino acid sequence described in SEQ ID NO:3; a nucleotide sequence encoding alkaline pullulanase possessing an amino acid sequence described in SEQ ID NO:4; recombinant DNAs containing these nucleotide sequences, and transformed microorganisms harboring the recombinant DNAs. The technique of the present invention enables mass production of alkaline pullulanase exhibiting both alkaline &agr;-amylase and alkaline pullulanase activity.
摘要翻译：本发明提供：编码碱基支链淀粉酶并表现出碱性α-淀粉酶和碱性支链淀粉酶活性的核苷酸序列; 编码具有SEQ ID NO：3所述的氨基酸序列的碱性α-淀粉酶的核苷酸序列; 编码具有SEQ ID NO：4所述的氨基酸序列的碱基支链淀粉酶的核苷酸序列; 含有这些核苷酸序列的重组DNA和携带重组DNA的转化微生物。本发明的技术能够大量生产表现出碱性α-淀粉酶和碱性支链淀粉酶活性的碱性支链淀粉酶。

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