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7 条记录 (耗时 0.026 秒)

    Process for producing peptides in E. coli
    1.
    发明授权
    Process for producing peptides in E. coli 失效
    在大肠杆菌中生产肽的方法

    公开(公告)号：US5670340A

    公开(公告)日：1997-09-23

    申请号：US352179

    申请日：1994-12-05

    申请人： Masayuki Yabuta ,  Yuji Suzuki ,  Kazuhiro Ohsuye ,  Takehiro Oshima ,  Seiko Onai ,  Koji Magota ,  Shoji Tanaka

    发明人： Masayuki Yabuta ,  Yuji Suzuki ,  Kazuhiro Ohsuye ,  Takehiro Oshima ,  Seiko Onai ,  Koji Magota ,  Shoji Tanaka

    IPC分类号： C12N15/09 ,  C07K14/58 ,  C07K14/585 ,  C12N9/38 ,  C12N15/16 ,  C12N15/62 ,  C12N15/67 ,  C12N15/70 ,  C12P21/02 ,  C12P21/06 ,  C12R1/19 ,  C12P21/00

    CPC分类号： C07K14/58 ,  C07K14/585 ,  C12N15/62 ,  C12N9/2471 ,  C12Y302/01023 ,  C07K2319/00 ,  C07K2319/23 ,  C07K2319/43 ,  C07K2319/50 ,  C07K2319/75 ,  C07K2319/90

    摘要： The present invention is a process to express a target peptide in a large amount and accumulate the target peptide in host cells in the form of inclusion bodies. The process comprises: A) culturing host cells transformed with a plasmid able to express a gene coding for a fusion protein represented in the formula A--L--B, wherein B is a target peptide, A is a protective peptide comprising a 90-210 amino acid fragment E. coli .beta.-galactosidase, and L is a linker peptide positioned between the C-terminus of the protective peptide and the N-terminus of the target peptide and selected so that when the fusion protein is treated by an enzyme or chemical substance, the target peptide is separated, and wherein the protective peptide and linker peptide are selected so that the isoelectric point of the fusion protein in between 4.9 and 6.9; B) obtaining an insoluble fraction comprising inclusion bodies by homogenization of th cultured transformed cells; C) solubilizing the fusion protein in the inclusion bodies by treatment of the insoluble fraction with a solubilizing agent; and, D) cleaving the peptide bond between the C-terminus of the linker peptide and the N-terminus of the target peptide of the solubilized fusion protein to release the target peptide from the other peptides followed by purification of the target peptide.

    摘要翻译： 本发明是大量表达目标肽并以包涵体的形式在宿主细胞中聚集靶肽的方法。 该方法包括：A）培养用能够表达编码式ALB中表示的融合蛋白的基因的质粒转化的宿主细胞，其中B是靶肽，A是包含90-210个氨基酸片段E的保护肽 大肠杆菌β-半乳糖苷酶，L是位于保护肽的C-末端和靶肽的N末端之间的连接肽，并且被选择使得当融合蛋白被酶或化学物质处理时，靶 分离肽，并且其中选择保护性肽和接头肽，使得融合蛋白的等电点在4.9和6.9之间; B）通过均质培养的转化细胞获得包含包含体的不溶性级分; C）通过用增溶剂处理不溶性级分将融合蛋白溶解在包涵体中; 并且D）切割接头肽的C末端和溶解的融合蛋白的靶肽的N末端之间的肽键，从其他肽释放靶肽，然后纯化目标肽。

    Use of a 97 amino acid leader sequence from the E. coli B-galactosidase gene for the production of hanp and hptc as fusion proteins
    2.
    发明授权
    Use of a 97 amino acid leader sequence from the E. coli B-galactosidase gene for the production of hanp and hptc as fusion proteins 失效
    使用大肠杆菌B-半乳糖苷酶基因的97个氨基酸的前导序列产生作为融合蛋白的hpp和hptc

    公开(公告)号：US4987070A

    公开(公告)日：1991-01-22

    申请号：US163548

    申请日：1988-03-03

    申请人： Koji Magota ,  Takehiro Oshima ,  Shoji Tanaka

    发明人： Koji Magota ,  Takehiro Oshima ,  Shoji Tanaka

    IPC分类号： C12N15/09 ,  C07K14/58 ,  C07K14/585 ,  C12N9/38 ,  C12N15/62 ,  C12N15/71 ,  C12P21/02 ,  C12R1/19

    CPC分类号： C07K14/58 ,  C07K14/585 ,  C12N15/62 ,  C12N15/71 ,  C12N9/2471 ,  C12Y302/01023 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/61 ,  C07K2319/75

    摘要： A process for the production of a physiologically active peptide (a target peptide) containing cysteine residue, comprising the steps of:(1) culturing Escherichia coli transformed with a plasmid capable of expressing a fusion protein under control of a promoter of E. coli origin or a promoter of a phage origin, wherein the fusion protein is represented by the following formula:A--L--B wherein B represents a target peptide containing cysteine residue; A represents a partner polypeptide consisting of 90 to 220 amino acid residue but not containing cysteine residue; and L represents a linker amino acid residue positioned between C-terminal of the partner polypeptide and N-terminal of the target peptide wherein the same amino acid as the linker amino acid is not present in the target peptide, and the linker amino acid is selected so that the peptide bond between the C-terminal of the linker amino acid and the N-terminal of the target peptide is claimed by a protease or the linker amino acid is selectively degraded by a chemical substance;(2) disrupting the cultured cells and obtaining an insoluble fraction containing the fusion protein;(3) solubilizing the fusion protein with a solubilizing agent, and treating the solubilizing fusion protein with the protease or the chemical substance to liberate the target peptide, and isolating the target peptide.

    Expression vector for human TNF
    3.
    发明授权
    Expression vector for human TNF 失效
    人TNF的表达载体

    公开(公告)号：US5059530A

    公开(公告)日：1991-10-22

    申请号：US402675

    申请日：1989-09-05

    申请人： Takehiro Oshima ,  Shoji Tanaka ,  Shigekazu Matsukura

    发明人： Takehiro Oshima ,  Shoji Tanaka ,  Shigekazu Matsukura

    IPC分类号： A61K38/00 ,  C07K14/525 ,  C12N1/21 ,  C12N15/70

    CPC分类号： C07K14/525 ,  C12N15/70 ,  A61K38/00

    摘要： The present invention provides a vector plasmid capable of efficient tumor necrosis factor (TNF) production, a process capable of efficient TNF production in a host transformed with said plasmid and a composition containing the TNF produced by said process.The novel plasmid of the present invention is characterized by having inserted therein a DNA fragment that has a phage-derived promoter region upstream of a structural gene for TNF and in which a DNA fragment containing an E. coli gene-derived transcription termination coding base sequence (terminator) is joined immediately downstream of a base sequence coding for the termination of translation of said structural gene.

    摘要翻译： 本发明提供能够有效的肿瘤坏死因子（TNF）产生的载体质粒，能够在用所述质粒转化的宿主中有效地产生TNF的方法和含有由所述方法产生的TNF的组合物。 本发明的新型质粒的特征在于，在其中插入有在TNF结构基因上游具有噬菌体衍生的启动子区域的DNA片段，其中含有大肠杆菌基因的转录终止编码碱基序列的DNA片段 （终止子）紧接在编码所述结构基因翻译终止的碱基序列的下游。

    DNA encoding a KEX2 endoprotease without a C-terminal hydrophobic region
    7.
    发明授权
    DNA encoding a KEX2 endoprotease without a C-terminal hydrophobic region 失效
    编码没有C末端疏水区的KEX2内切蛋白的DNA

    公开(公告)号：US5234830A

    公开(公告)日：1993-08-10

    申请号：US916627

    申请日：1992-07-22

    申请人： Takehiro Oshima ,  Kensaku Mizuno

    发明人： Takehiro Oshima ,  Kensaku Mizuno

    IPC分类号： C12N9/60

    CPC分类号： C12N9/60 ,  C07K2319/00

    摘要： A KEX2 endoprotease produced by a recombinant DNA technique, a KEX2 endoprotease shortened at the C-terminal of a native enzyme and still containing a C-terminal hydrophobic region, a soluble KEX2 endoprotease without a C-terminal hydrophobic region; DNA's coding for the above-mentioned enzymes; expression plasmids containing the DNA; hosts transformed with the plasmid; a process for production of the above-mentioned enzymes using the transformed host; and a process for production of biologically active polypeptide using the above-mentioned enzyme.

    摘要翻译： 通过重组DNA技术产生的KEX2内切蛋白酶，在天然酶的C末端缩短且仍含有C末端疏水区域的KEX2内切蛋白酶，不具有C末端疏水区域的可溶性KEX2内切蛋白酶; DNA编码上述酶; 含有DNA的表达质粒; 用质粒转化的宿主; 使用转化的宿主生产上述酶的方法; 以及使用上述酶生产生物活性多肽的方法。