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    NOVEL RECOMBINED PLASMID PBLAK1
    31.
    发明专利
    NOVEL RECOMBINED PLASMID PBLAK1 失效

    公开(公告)号：JPS63245679A

    公开(公告)日：1988-10-12

    申请号：JP7937787

    申请日：1987-03-31

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： IWAKURA MASAHIRO ,  TSUDA KEISHIRO

    IPC: C12N15/09 ,  C07K7/18 ,  C12N1/20 ,  C12N1/21 ,  C12N9/06 ,  C12P21/02 ,  C12R1/19

    Abstract: PURPOSE:To make up a novel recombined plasmid pBLAK1 including a gene coding a fused protein from dihydrofolate reductase and bradykinin, and trans form E. coli with the plasmid to produce bradykinin. CONSTITUTION:The recombined plasmid pBLAK1 is stably copied in E. coli and gives the E. coli, the host, resistance to trimethoprim and ampicillin. The gene giving resistance to trimethoprim is recombined by modifying the 3'-termi nal of the dihydrofolate gene from Bacillus subtilis to code the fused protein from dihydrofolate reductase-bradykinin with 4474 pairs of bases. The recombinant plasmid pBLAK1 is introduced into E. coli C600 strain and the transformed strain is deposited as FERM P-9300.

    NOVEL RECOMBINANT PLASMID CONTAINING LEUCINE ENKEPHALIN GENE
    32.
    发明专利
    NOVEL RECOMBINANT PLASMID CONTAINING LEUCINE ENKEPHALIN GENE 失效

    公开(公告)号：JPS61260885A

    公开(公告)日：1986-11-19

    申请号：JP10087885

    申请日：1985-05-13

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： IWAKURA MASAHIRO ,  FURUSAWA KIYOTAKA ,  KOKUBU TOMOKUNI ,  TSUDA KEISHIRO

    IPC: C12N15/09 ,  C07K14/70 ,  C12N1/20 ,  C12P21/02 ,  C12R1/19

    Abstract: PURPOSE:To construct a specific recombinant plasmid having a specific DNA sequence, by making it possible to cut out a DNA sequence coding leucine enkephalin consisting of 28 base pairs by cleavage with a restriction enzyme EcoRI. CONSTITUTION:A novel recombinant plasmid pLEG1 obtained by carrying out sorting out of a plasmid vector and chemical synthesis of a DNA coding leucine enkephalin, and passing the synthesized DNA through processes of integration into a plasmid vector, introduction of the resultant recombinant plasmid into Escherichia coli K12C600 strain, separation of microbial cells having the recombinant plasmid and separation of the plasmid. DNAs expressed by formulas I and II synthesized by the phosphoramidite solid phase synthesis method may be used as the DNA coding the leucine enkephalin.

    NEW SYNTHESIS OF PEPTIDE
    33.
    发明专利
    NEW SYNTHESIS OF PEPTIDE 失效

    公开(公告)号：JPH1045798A

    公开(公告)日：1998-02-17

    申请号：JP20377196

    申请日：1996-08-01

    Applicant: AGENCY IND SCIENCE TECHN ,  EISAI CO LTD

    Inventor： IWAKURA MASAHIRO ,  TAKENAWA TATSUYUKI ,  YOSHINARI KOICHI ,  ODA YOSHIYA ,  ISHIHAMA YASUSHI

    IPC: C07K1/02 ,  C07K7/06

    Abstract: PROBLEM TO BE SOLVED: To provide a method for synthesizing a peptide in block units by directly bonding the mutual peptides. SOLUTION: A peptide chain having a cyanocysteine residue R1 -CO-NH-C (CH2 -SCN)-CO-NH-R2 (R1 and R2 denote each an optional amino acid sequence) is reacted with another peptide chain having the free amino terminal NH2 -R3 (R3 denotes an optional amino acid sequence). Thereby, amino groups in the peptide chain make a nucleophilic attack on carbonyl carbons on the peptide bond to initiate bonding reaction between peptide bonds. As a result, a peptide chain having a sequence of R1 -CO-NH-R3 is synthesized.

    SEPARATION AND PURIFICATION OF RECOMBINANT PROTEIN
    37.
    发明专利
    SEPARATION AND PURIFICATION OF RECOMBINANT PROTEIN 失效

    公开(公告)号：JPH02255697A

    公开(公告)日：1990-10-16

    申请号：JP7613489

    申请日：1989-03-28

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： IWAKURA MASAHIRO

    IPC: C12N15/09 ,  C07K1/14 ,  C07K1/16 ,  C07K1/20 ,  C07K1/22 ,  C07K14/00 ,  C07K19/00 ,  C12N15/62 ,  C12P21/00 ,  C12P21/02 ,  C12R1/19

    Abstract: PURPOSE:To solubilize, highly purify and homogenize a dihydrofolic acid- reducing enzyme-fused protein regenerated and accumulated in the bodies of Escherichia coli as an insoluble protein and to activate the enzymatic portion of the protein by employing a specific means. CONSTITUTION:A fused protein prepared by combining a different kind of protein with the carboxyl terminal group of the dihydrofolic acid-reducing enzyme of Escherichia coli is accumulated in the cells of the Escherichia coli as an insoluble protein by the expression of a gene coding the fused protein. The bodies of the Escherichia coli expressing and producing the insoluble protein are ground and centrifuged, and the precipitated fraction is solubilized with acetic acid. The solubilized fused protein is applied to a reverse phase liquid high performance chromatography to purify in a high degree. The Escherichia coli includes Escherichia coli (FERMBP-2149) containing a recombined plasmid pSG1-12.

    BRADYKININ
    39.
    发明专利
    BRADYKININ 失效

    公开(公告)号：JPH01252286A

    公开(公告)日：1989-10-06

    申请号：JP7967788

    申请日：1988-03-31

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： IWAKURA MASAHIRO ,  OHASHI SHINICHI ,  SAKAI TSUKASA ,  TANAKA YOSHIO

    IPC: C12N15/09 ,  C07K1/12 ,  C07K1/16 ,  C07K1/22 ,  C07K7/18 ,  C07K14/00 ,  C07K19/00 ,  C12N1/20 ,  C12N1/21 ,  C12N9/06 ,  C12P21/02 ,  C12R1/19

    Abstract: PURPOSE:To obtain a large amount of dihydrofolic acid reductase-bradykinin fused protein, by manifesting a fused gene in which a bradykinin gene is included in a plasmid vector pTP 70-1 with a colon bacillus. CONSTITUTION:A newly recombined plasmid pBK 1-11, which is replicated in stable with a colon bacillus, imparting resistance to trimethoprim and resistance to ampicillin to a colon bacillus as a host, having a size of 4645 basic pairs and having a bonded structure of chemically synthesized DNA of 37 basic pairs containing a conformation coding bradykinin at BamHI site of pTP 70-1, is produced. A colon bacillus introduced of pBK 1-11 (FERMBP-1817) produces a large amount of dihydrofolic acid reductase-bradykinin fused protein.

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