公开(公告)号：US5827710A

公开(公告)日：1998-10-27

申请号：US869506

申请日：1997-06-05

Applicant: Kohji Uchida ,  Hirokazu Matsukawa ,  Tuyosi Fujita ,  Yushi Matuo

Inventor： Kohji Uchida ,  Hirokazu Matsukawa ,  Tuyosi Fujita ,  Yushi Matuo

IPC: C12N15/09 ,  C07H21/04 ,  C12N1/16 ,  C12N9/04 ,  C12Q1/52 ,  C12R1/865 ,  C12N1/14 ,  C12N15/00

CPC classification number: C12N9/0006 ,  C12Y101/01027 ,  C12Y101/01028

Abstract: The present invention provides a gene encoding the type B subunit protein of avian lactate dehydrogenase. The gene encoding the type B subunit protein of avian lactate dehydrogenase of the present invention is obtained by plaque hybridization or the like from a cDNA library derived from avian heart muscle and encodes the type B subunit protein of avian lactate dehydrogenase having the sequences shown as SEQ ID NO. 1 or 2.

Abstract translation: 本发明提供编码禽类乳酸脱氢酶的B型亚基蛋白的基因。 编码本发明禽类乳酸脱氢酶B型亚基蛋白的基因是通过来自禽心肌的cDNA文库的斑块杂交等获得的，并编码具有SEQ所示序列的禽类乳酸脱氢酶的B亚基蛋白 证件号码。 1或2。

公开(公告)号：US6057141A

公开(公告)日：2000-05-02

申请号：US128967

申请日：1998-08-04

Applicant: Kohji Uchida ,  Hirokazu Matsukawa ,  Tuyosi Fujita ,  Yushi Matuo

Inventor： Kohji Uchida ,  Hirokazu Matsukawa ,  Tuyosi Fujita ,  Yushi Matuo

IPC: C12N9/04 ,  A23J1/00 ,  C12N5/00

CPC classification number: C12N9/0006 ,  C12Y101/01027 ,  C12Y101/01028

Abstract: The present invention provides a gene encoding the type B subunit protein of avian lactate dehydrogenase. The gene encoding the type B subunit protein of avian lactate dehydrogenase of the present invention is obtained by plaque hybridization or the like from a cDNA library derived from avian heart muscle and encodes the type B subunit protein of avian lactate dehydrogenase having an amino acid sequence shown as SEQ ID NO. 2.

Abstract translation: 本发明提供编码禽类乳酸脱氢酶的B型亚基蛋白的基因。 本发明的编码禽类乳酸脱氢酶B型亚单位蛋白的基因是通过来自禽心肌的cDNA文库的斑块杂交等获得的，并编码具有氨基酸序列的禽类乳酸脱氢酶的B亚基蛋白 如SEQ ID NO。 2。

公开(公告)号：US5952211A

公开(公告)日：1999-09-14

申请号：US941647

申请日：1997-09-30

Applicant: Atsuo Nakamura ,  Toshio Tanaka ,  Yushi Matuo ,  Sumio Tanase ,  Masahiko Funatsu ,  Akira Eto

Inventor： Atsuo Nakamura ,  Toshio Tanaka ,  Yushi Matuo ,  Sumio Tanase ,  Masahiko Funatsu ,  Akira Eto

IPC: G01N33/53 ,  C07H21/04 ,  C07K14/47 ,  C12N1/21 ,  C12N9/10 ,  C12N15/09 ,  C12P21/02 ,  C12Q1/68 ,  C12R1/19 ,  G01N33/573

CPC classification number: C12N9/1096

Abstract: The invention relates to a mutated human ALT (alanine aminotransferase) gene improving the DNA sequence encoding the N-terminal region of human ALT, without causing amino acid substitution, while, at the same time, adding restriction endonuclease sites to the upstream and downstream sites of the gene. By cultivation of E. coli transformant carrying a recombinant plasmid, which was constructed by ligation of the mutated human ALT gene into a vector, an active human ALT can be produced efficiently.

Abstract translation: 本发明涉及改善编码人ALT的N末端区域的DNA序列而不引起氨基酸取代的突变人ALT（丙氨酸氨基转移酶）基因，同时向上游和下游位点添加限制性内切核酸酶位点 的基因。 通过将通过将突变的人ALT基因连接到载体中构建的携带重组质粒的大肠杆菌转化体的培养，可以有效地生产活性人ALT。

公开(公告)号：US5888766A

公开(公告)日：1999-03-30

申请号：US536941

申请日：1995-09-29

Applicant: Toshihiro Ishizuka ,  Yasunori Kihira ,  Toshio Tanaka ,  Yushi Matuo

Inventor： Toshihiro Ishizuka ,  Yasunori Kihira ,  Toshio Tanaka ,  Yushi Matuo

IPC: C12N15/09 ,  C07H21/04 ,  C07K14/805 ,  C12N1/21 ,  C12P21/02 ,  C12R1/19 ,  C12P21/06 ,  C07H17/00 ,  C12N1/20

CPC classification number: C07K14/805

Abstract: The present invention relates to a method for producing human myoglobin by inserting a DNA sequence comprising a human myoglobin gene into a tryptophan promoter-containing vector, followed by making E. coli with the thus-obtained expression vector express a holo-type human myoglobin. Using this method, it is possible to directly and efficiently produce a heme-structured, holo-type human myoglobin by gene recombination technology.

Abstract translation: 本发明涉及一种通过将含有人肌红蛋白基因的DNA序列插入含色氨酸启动子的载体中，然后用由此获得的表达载体表达全息型人肌红蛋白的大肠杆菌来生产人肌红蛋白的方法。 使用这种方法，可以通过基因重组技术直接有效地产生血红素结构的全息型人肌红蛋白。

公开(公告)号：US06265159B1

公开(公告)日：2001-07-24

申请号：US08998712

申请日：1997-12-29

Applicant: Yoshinobu Sugino ,  Masayuki Morita ,  Yushi Matuo ,  Kohji Uchida

Inventor： Yoshinobu Sugino ,  Masayuki Morita ,  Yushi Matuo ,  Kohji Uchida

IPC: C12Q168

CPC classification number: C12N15/00

Abstract: The present invention provides a method for producing nested deletions in vitro in desired DNA, comprising: 1) preparing a vector containing a DNA fragment in which nested deletions are to be generated and the terminal repeat of a transposon; 2) incubating the vector in vitro with transposase and a DNA replication system; 3) obtaining the vector incorporating the DNA fragment with nested deletions as a reaction product; and optionally, 4) transforming a host cell with the reaction product and growing it; and 5) recovering a vector incorporating said DNA fragment with nested deletions from the grown host cell.

Abstract translation: 本发明提供了一种用于在所需DNA中体外产生嵌套缺失的方法，包括：1）制备含有其中产生嵌套缺失的DNA片段和转座子末端重复的载体; 2）体外培养载体 具有转座酶和DNA复制体系; 3）获得含有嵌合缺失的DNA片段作为反应产物的载体; 和任选地，4）用反应产物转化宿主细胞并使其生长; 和5）从生长的宿主细胞中回收含有嵌套缺失的所述DNA片段的载体。

公开(公告)号：US06387683B1

公开(公告)日：2002-05-14

申请号：US09368588

申请日：1999-08-05

Applicant: Nobuyoshi Ishii ,  Yasuo Suzuki ,  Kohji Uchida ,  Yushi Matuo ,  Hideo Tanaka

Inventor： Nobuyoshi Ishii ,  Yasuo Suzuki ,  Kohji Uchida ,  Yushi Matuo ,  Hideo Tanaka

IPC: C12N990

CPC classification number: C12N9/90 ,  Y10S435/942

Abstract: The present invention provides a process for producing biologically active recombinant yeast protein disulfide isomerase, which includes the steps of: a) deleting, substituting, or adding one or more bases in a region encoding an endoplasmic reticulum retention signal in a gene encoding protein disulfide isomerase of yeast to modify the gene so as not to encode part or all of the endoplasmic reticulum retention signal; b) incorporating the modified gene into an expression vector; c) transforming host cells with the expression vector; and d) culturing the host cells transformed with the expression vector in a culture medium kept at a nearly neutral pH, thereby causing protein disulfide isomerase to be secreted in an active state outside the host cells. According to the process of the invention, recombinant yeast protein disulfide isomerase can be secreted in a large amount in a biologically active state into a culture medium, and can be collected by a simple purification method.

Abstract translation: 本发明提供生产生物活性重组酵母蛋白二硫键异构酶的方法，其包括以下步骤：a）在编码蛋白质二硫键异构酶的基因的编码内质网保持信号的区域中缺失，取代或添加一个或多个碱基 的酵母修饰基因，以免编码部分或全部内质网保持信号; b）将修饰的基因并入表达载体中; c）用表达载体转化宿主细胞; 和d）在保持接近中性pH的培养基中培养用表达载体转化的宿主细胞，从而使蛋白质二硫键异构酶以宿主细胞外的活性状态分泌。 根据本发明的方法，可以将重组酵母蛋白二硫键异构酶以大量的生物活性状态分泌到培养基中，并且可以通过简单的纯化方法收集。