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    Extracellularly secreted nuclease
    2.
    发明授权
    Extracellularly secreted nuclease 有权
    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。

    Plasmid vector
    4.
    发明授权
    Plasmid vector 有权
    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）。

    METHOD FOR DETACHMENT AND PREPARATION OF LIVING MICROORGANISMS
    5.
    发明申请
    METHOD FOR DETACHMENT AND PREPARATION OF LIVING MICROORGANISMS 有权
    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。

    NOVEL EXTRACELLULARLY SECRETED NUCLEASE
    6.
    发明申请
    NOVEL EXTRACELLULARLY SECRETED NUCLEASE 有权
    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。

    PLASMID VECTOR
    7.
    发明申请
    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）。

    METHODS FOR STABLY RETAINING FOREIGN GENES IN CELLS
    9.
    发明申请
    METHODS FOR STABLY RETAINING FOREIGN GENES IN CELLS 审中-公开
    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。