公开(公告)号：US5409815A

公开(公告)日：1995-04-25

申请号：US311556

申请日：1989-02-16

申请人： Yukimitsu Nakagawa ,  Naofumi Hayasuke ,  Atsushi Masaki ,  Yutaka Ishida ,  Kohji Murakami ,  Ken Okabayashi ,  Kiyoshi Tsutsui ,  Hitoshi Minamino ,  Kazuo Ikegaya ,  Kaoru Kobayashi ,  Haruhide Kawabe ,  Sadao Ueda

发明人： Yukimitsu Nakagawa ,  Naofumi Hayasuke ,  Atsushi Masaki ,  Yutaka Ishida ,  Kohji Murakami ,  Ken Okabayashi ,  Kiyoshi Tsutsui ,  Hitoshi Minamino ,  Kazuo Ikegaya ,  Kaoru Kobayashi ,  Haruhide Kawabe ,  Sadao Ueda

IPC分类号： C12N15/09 ,  C07K7/06 ,  C07K7/08 ,  C07K14/00 ,  C07K14/705 ,  C07K14/765 ,  C07K19/00 ,  C12N1/16 ,  C12N15/62 ,  C12N15/81 ,  C12P21/02 ,  C12R1/865 ,  C12P21/06 ,  C07H21/04

CPC分类号： C12N15/81 ,  C07K14/765 ,  C12N15/625 ,  C07K2319/02 ,  C07K2319/55 ,  C07K2319/61

摘要： Novel signal peptides capable of functioning in yeasts have the amino acid sequence:Met-A.sub.1 -A.sub.2 -X-B-C-D-E-Fwherein A.sub.1 is a peptide chain composed of 1-3 amino acids each selected from the group consisting of Arg, Ser, Lys and His, A.sub.2 is a peptide chain composed of 1-3 amino acids, X is a peptide chain composed of 8-10 hydrophobic amino acids, B is Pro or Ser, C is Gly or Pro, D Cys, Ala, Leu, Ser, Thr and Val, E is Trp or Gln and F is Ala or Gly. DNA sequences coding for these signal peptides can be used for the secretory expression of heterologous proteins in yeasts.

摘要翻译： 能够在酵母中起作用的新型信号肽具有氨基酸序列：Met-A1-A2-XBCDEF，其中A1是由1-3个氨基酸组成的肽链，每个氨基酸选自Arg，Ser，Lys和His，A2 是由1-3个氨基酸组成的肽链，X是由8-10个疏水性氨基酸组成的肽链，B是Pro或Ser，C是Gly或Pro，D Cys，Ala，Leu，Ser，Thr和Val ，E是Trp或Gln，F是Ala或Gly。 编码这些信号肽的DNA序列可用于酵母中异源蛋白质的分泌表达。

公开(公告)号：US4945046A

公开(公告)日：1990-07-31

申请号：US57143

申请日：1987-06-03

申请人： Hajime Horii ,  Haruhide Kawabe ,  Hirofumi Arimura ,  Hiromichi Mukai ,  Kaoru Kobayashi ,  Muneo Tsujikawa ,  Masayuki Nishida ,  Tadakazu Suyama

发明人： Hajime Horii ,  Haruhide Kawabe ,  Hirofumi Arimura ,  Hiromichi Mukai ,  Kaoru Kobayashi ,  Muneo Tsujikawa ,  Masayuki Nishida ,  Tadakazu Suyama

IPC分类号： C07K14/02 ,  C12N15/81

CPC分类号： C07K14/005 ,  C12N15/81 ,  C12N2730/10122

摘要： A method of producing a heterologous protein in yeast is disclosed, which comprises transforming a yeast Saccharomyces cerevisiae with recombinant DNA comprising a promoter selected from the group consisting of a yeast promoter or a hybrid promoter derived from a yeast promoter, particularly a hybrid promoter containing the enhancer region of SV40 virus, and a gene coding for a heterologous protein such as HBsAg and Pre S-HBsAg, particularly full length Pre S-HBsAg culturing, the resulting transformed cells to express the gene coding the heterologous protein, and isolating the heterologous protein from the cultured medium. GAP-DH and PH05 promoters and miniaturized ones can be used as the yeast promoter. The expression of the full length Pre S-HBsAg is carried out in the same expression system as that for 2nd or 3rd Pre S-ABsAg.

摘要翻译： 公开了一种在酵母中产生异源蛋白质的方法，其包括用包含启动子的重组DNA转化酵母酿酒酵母，所述启动子选自酵母启动子或源自酵母启动子的杂合启动子，特别是含有 SV40病毒的增强子区和编码异源蛋白如HBsAg和Pre S-HBsAg的基因，特别是全长Pre S-HBsAg培养，所得转化细胞表达编码异源蛋白的基因，并分离异源蛋白 从培养基中。 GAP-DH和PH05启动子和小型化可用作酵母启动子。 全长前S-HBsAg的表达在与第2或第3前S-ABsAg相同的表达系统中进行。

公开(公告)号：US5756313A

公开(公告)日：1998-05-26

申请号：US138384

申请日：1993-10-20

申请人： Ken Okabayashi ,  Hideyuki Oi ,  Kazumoto Hirabayashi ,  Masami Miura ,  Miho Shimizu ,  Haruhide Kawabe

发明人： Ken Okabayashi ,  Hideyuki Oi ,  Kazumoto Hirabayashi ,  Masami Miura ,  Miho Shimizu ,  Haruhide Kawabe

IPC分类号： C12N1/19 ,  C07K14/765 ,  C12N9/26 ,  C12N15/09 ,  C12N15/14 ,  C12N15/81 ,  C12N15/90 ,  C12P21/00 ,  C12P21/02 ,  C12R1/865 ,  C12N15/00

CPC分类号： C07K14/765 ,  C12N15/81 ,  C12N15/90 ,  C12N9/2408 ,  C07K2319/02

摘要： A plasmid for a yeast host, the plasmid comprising in sequence: (1) a yeast derived promoter, (2) an albumin-encoding region placed under control of the yeast-derived promoter, (3) a transcription terminator and (4) a sequence homologous to a part of the yeast host chromosomal sequence such that the plasmid is capable of being integrated into the yeast host cell chromosome and wherein the plasmid is incapable of autonomous replication in yeast cells; a yeast host transformed with the above plasmid; a method of producing a yeast transformant which comprises transforming a yeast host by integrating at least two different plasmids each having a sequence homologous to a part of the host yeast chromosomal sequence one at a time into the chromosome of the yeast host cell; and a method of producing albumin which comprises cultivating the above transformant and recovering the thus-produced albumin.

摘要翻译： 一种用于酵母宿主的质粒，所述质粒按顺序包含：（1）酵母衍生的启动子，（2）置于酵母衍生的启动子控制下的白蛋白编码区，（3）转录终止子和（4） 序列与酵母宿主染色体序列的一部分同源，使得质粒能够整合到酵母宿主细胞染色体中，并且其中质粒不能在酵母细胞中自主复制; 用上述质粒转化的酵母宿主; 生产酵母转化体的方法，其包括通过将至少两种与宿主酵母染色体部分的序列同源的不同质粒整合到酵母宿主细胞的染色体中来转化酵母宿主; 以及生产白蛋白的方法，其包括培养上述转化体并回收由此产生的白蛋白。

公开(公告)号：US5021339A

公开(公告)日：1991-06-04

申请号：US397347

申请日：1989-08-24

申请人： Hiromichi Mukai ,  Hajime Horii ,  Muneo Tsujikawa ,  Haruhide Kawabe ,  Hirofumi Arimura ,  Tadakazu Suyama

发明人： Hiromichi Mukai ,  Hajime Horii ,  Muneo Tsujikawa ,  Haruhide Kawabe ,  Hirofumi Arimura ,  Tadakazu Suyama

IPC分类号： C07K14/02 ,  C12N15/81

CPC分类号： C07K14/005 ,  C12N15/81 ,  C12N2730/10122

摘要： A DNA sequence of the yeast glyceraldehyde-3-phosphate dehydrogenase (GAP-DH) promoter and a process for preparing a heterologous protein utilizing said DNA sequence as a promoter are disclosed, said GAP-DH promoter comprising a region upstream of an initiator codon of the GAP-DH protein up to -164 bp as a minimum unit. Since the GAP-DH promoter is small in size, a physiologically active substance can be expressed in yeasts effectively and recombination of DNA can be simplified.

摘要翻译： 公开了酵母甘油醛-3-磷酸脱氢酶（GAP-DH）启动子的DNA序列和利用所述DNA序列作为启动子制备异源蛋白质的方法，所述GAP-DH启动子包含上游的起始密码子 最高达-164 bp的GAP-DH蛋白作为最小单位。 由于GAP-DH启动子的大小小，所以能够有效地在酵母中表达生理活性物质，可以简化DNA的重组。

公开(公告)号：US4757021A

公开(公告)日：1988-07-12

申请号：US795144

申请日：1985-11-05

申请人： Haruhide Kawabe ,  Kazumoto Hirabayashi ,  Hazime Horii ,  Hirofumi Arimura ,  Masayuki Nishida ,  Tadakazu Suyama

发明人： Haruhide Kawabe ,  Kazumoto Hirabayashi ,  Hazime Horii ,  Hirofumi Arimura ,  Masayuki Nishida ,  Tadakazu Suyama

IPC分类号： C12N15/09 ,  C07K14/02 ,  C12N1/16 ,  C12N15/68 ,  C12N15/81 ,  C12P21/00 ,  C12R1/865 ,  C12N1/18 ,  C12N1/00 ,  C12N15/00

CPC分类号： C07K14/005 ,  C12N15/68 ,  C12N15/81 ,  C12N2730/10122

摘要： A recombinant plasmid wherein at least two DNA fragments containing a Baker gene and a Charlie gene of the yeast 2 .mu.m circular plasmid are incorporated into a plasmid containing genes encoding a physiologically active substance is disclosed. Also, a strain transformed by the recombinant plasmid is disclosed.

摘要翻译： 公开了将包含酵母2μm圆形质粒的Baker基因和Charlie基因的至少两个DNA片段并入包含编码生理活性物质的基因的质粒中的重组质粒。 此外，公开了由重组质粒转化的菌株。

公开(公告)号：US5610036A

公开(公告)日：1997-03-11

申请号：US288899

申请日：1994-08-10

申请人： Masami Miura ,  Yutaka Ishida ,  Hideyuki Oi ,  Koji Murakami ,  Yukimitsu Nakagawa ,  Haruhide Kawabe

发明人： Masami Miura ,  Yutaka Ishida ,  Hideyuki Oi ,  Koji Murakami ,  Yukimitsu Nakagawa ,  Haruhide Kawabe

IPC分类号： C07K14/765 ,  C12N9/26 ,  C12N15/14 ,  C12N15/81 ,  C07H21/04 ,  C12N1/16 ,  C12N1/19 ,  C12P21/02

CPC分类号： C12N9/2408 ,  C07K14/765 ,  C12N15/815

摘要： A mutant AOX2 (alcohol oxidase 2) promoter derived from the natural AOX2 promoter by base sequence deletion, insertion, or substitution is disclosed. A microbial strain carrying a gene under the control of such a mutant AOX2 promoter can be obtained by growing in a methanol-containing medium a strain incapable of producing AOX encoded by the AOX1 gene but carrying the AOX2 gene under the control of the natural AOX2 promoter. A heterologous protein may be produced by cultivating the mutant strain with the desired heterologous protein gene incorporated downstream from the mutant AOX2 promoter. A plasmid carrying the mutant AOX2 promoter is also provided.

摘要翻译： 公开了通过碱基序列缺失，插入或取代衍生自天然AOX2启动子的突变体AOX2（醇氧化酶2）启动子。 携带这种突变体AOX2启动子控制下的基因的微生物菌株可以通过在含甲醇培养基中生长不能产生由AOX1基因编码的AOX但携带AOX2基因在天然AOX2启动子控制下的菌株 。 可以通过用突变AOX2启动子下游掺入的所需异源蛋白质基因培养突变菌株来产生异源蛋白质。 还提供携带突变型AOX2启动子的质粒。

公开(公告)号：US5643792A

公开(公告)日：1997-07-01

申请号：US181242

申请日：1994-01-13

申请人： Ken Okabayashi ,  Takao Ohmura ,  Kazumasa Yokoyama ,  Haruhide Kawabe

发明人： Ken Okabayashi ,  Takao Ohmura ,  Kazumasa Yokoyama ,  Haruhide Kawabe

IPC分类号： C12N1/20 ,  C07K14/765 ,  C12N1/15 ,  C12N1/21 ,  C12N9/04 ,  C12N9/64 ,  C12N15/00 ,  C12N15/01 ,  C12N15/52 ,  C12N15/74 ,  C12N15/78 ,  C12N15/81 ,  C12P21/00 ,  C12R1/84 ,  C12N15/11

CPC分类号： C12N9/0006 ,  C07K14/765 ,  C12N15/52 ,  C12N15/74 ,  C12N15/78 ,  C12N15/815 ,  C12Y101/03013

摘要： A methylotrophic and glucotrophic mutant strain capable of producing a heterologous protein and a method for producing a heterologous protein, comprising culture of the mutant strain. The mutant strain of the present invention can be grown in a medium containing both methanol and glucose, with the effect that the growth of the strain and production of a heterologous protein proceed at the same time. Accordingly, a heterologous protein can be produced in a large amount in a short time.

摘要翻译： 能够产生异源蛋白质的甲基营养型和葡萄糖营养型突变菌株和产生异源蛋白质的方法，包括突变菌株的培养。 本发明的突变菌株可以在含有甲醇和葡萄糖的培养基中生长，其效果是菌株的生长和异源蛋白质的产生同时进行。 因此，可以在短时间内大量生产异源蛋白质。

公开(公告)号：US5503993A

公开(公告)日：1996-04-02

申请号：US445783

申请日：1995-05-22

申请人： Naofumi Hayasuke ,  Yukimitsu Nakagawa ,  Yutaka Ishida ,  Ken Okabayashi ,  Kohji Murakami ,  Kiyoshi Tsutsui ,  Kazuo Ikegaya ,  Hitoshi Minamino ,  Sadao Ueda ,  Haruhide Kawabe ,  Hirofumi Arimura ,  Atusi Masaki

发明人： Naofumi Hayasuke ,  Yukimitsu Nakagawa ,  Yutaka Ishida ,  Ken Okabayashi ,  Kohji Murakami ,  Kiyoshi Tsutsui ,  Kazuo Ikegaya ,  Hitoshi Minamino ,  Sadao Ueda ,  Haruhide Kawabe ,  Hirofumi Arimura ,  Atusi Masaki

IPC分类号： C12N1/19 ,  C07K14/765 ,  C12N15/09 ,  C12N15/81 ,  C12P21/02 ,  C12R1/865 ,  C12N15/14

CPC分类号： C07K14/765 ,  C12N15/81 ,  C07K2319/02

摘要： A method for preparing foreign protein in yeast using an expression recombinant DNA comprising DNA encoding the serum albumin signal peptide adjacent to DNA encoding the foreign protein is disclosed.

摘要翻译： 公开了一种使用包含编码与编码外源蛋白的DNA相邻的血清白蛋白信号肽的DNA的表达重组DNA在酵母中制备外源蛋白的方法。

公开(公告)号：US5254470A

公开(公告)日：1993-10-19

申请号：US672304

申请日：1991-03-20

申请人： Seiji Murakami ,  Hiroki Tatsumi ,  Yoshihiro Ogawa ,  Eiichi Nakano ,  Hiroshi Motai ,  Shigetoshi Sugio ,  Atsushi Masaki ,  Yutaka Ishida ,  Kohji Murakami ,  Haruhide Kawabe ,  Hirofumi Arimura

发明人： Seiji Murakami ,  Hiroki Tatsumi ,  Yoshihiro Ogawa ,  Eiichi Nakano ,  Hiroshi Motai ,  Shigetoshi Sugio ,  Atsushi Masaki ,  Yutaka Ishida ,  Kohji Murakami ,  Haruhide Kawabe ,  Hirofumi Arimura

IPC分类号： C12N15/09 ,  C12N9/62 ,  C12N15/51 ,  C12R1/69

CPC分类号： C12N9/62

摘要： Alkaline protease which has leucine or isoleucine in the place of valine at the amino acid number 40 of wild type alkaline protease, a gene encoding the amino acid sequence of alkaline protease which has the substitution as described above, a recombinant DNA comprising said gene, a method of producing the above described alkaline protease, and a DNA fragment used for the expression of a gene.

摘要翻译： 在野生型碱性蛋白酶的氨基酸数40处含有亮氨酸或异亮氨酸代替缬氨酸的碱性蛋白酶，编码如上所述取代的碱性蛋白酶的氨基酸序列的基因，包含所述基因的重组DNA， 制备上述碱性蛋白酶的方法和用于表达基因的DNA片段。

公开(公告)号：US5098840A

公开(公告)日：1992-03-24

申请号：US525011

申请日：1990-05-18

申请人： Shunji Kasai ,  Ryuji Hiramatsu ,  Shusei Uno ,  Masanori Nagai ,  Hirofumi Arimura ,  Toshizumi Tanabe ,  Yasuo Amatsuji ,  Masaaki Hirose ,  Masanori Morita ,  Haruhide Kawabe

发明人： Shunji Kasai ,  Ryuji Hiramatsu ,  Shusei Uno ,  Masanori Nagai ,  Hirofumi Arimura ,  Toshizumi Tanabe ,  Yasuo Amatsuji ,  Masaaki Hirose ,  Masanori Morita ,  Haruhide Kawabe

IPC分类号： A61K38/00 ,  C12N9/72 ,  C12N15/58

CPC分类号： C12N9/6462 ,  C12Y304/21073 ,  A61K38/00

摘要： A human prourokinase mutant in which the entire or a partial epidermal growth factor domain of human prourokinase is deleted or a partial epidermal growth factor domain of human prourokinase is replaced by one or more different amino acid residues, said mutant having a longer blood half-life than naturally occurring human prourokinase while retaining prourokinase enzymatic activity. In this human prourokinase mutant the region selected from the group consisting of: (a) from asparagine (10) to cysteine (42); (b) from asparagine (10) to aspartic acid (45); and (c) from asparagine (10) to threonine (49) is missing.

摘要翻译： 其中缺少人原尿激酶的全部或部分表皮生长因子结构域的人prourokinase突变体或人原尿激酶的部分表皮生长因子结构域被一个或多个不同的氨基酸残基替代，所述突变体具有较长的血液半衰期 而不是天然存在的人类prourokinase，同时保留prourokinase酶活性。 在该人类prourokinase突变体中，选自：（a）天冬酰胺（10）至半胱氨酸（42）的区域; （b）从天冬酰胺（10）到天冬氨酸（45）; 和（c）从天冬酰胺（10）到苏氨酸（49）缺失。