公开(公告)号：JPS63102696A

公开(公告)日：1988-05-07

申请号：JP24926086

申请日：1986-10-20

Applicant: AGENCY IND SCIENCE TECHN

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

IPC: C12N15/09 ,  C07K1/16 ,  C07K14/575 ,  C07K14/655 ,  C07K14/70 ,  C07K19/00 ,  C12N9/06 ,  C12N15/00 ,  C12P21/00 ,  C12P21/02 ,  C12R1/19

Abstract: PURPOSE:To obtain a dihydrofolate reductase-leucine enkephalin fusion protein which is protein produced by Escherichia coli integrated with a specific plasmid and a raw material for leucine enkephalin having nonhabitual morphine-like analgesic action. CONSTITUTION:Escherichia coli integrated with plasmid pBSFOLEK1 is cultivated to produce dihydrofolate reductase-leucine enkephalin fusion protein having an amino acid sequence shown by the fig. The cell is separated from the culture solution, removed, the culture solution is subjected to ion exchange chromatography and then the aimed fusion protein is separated and purified by using dihydrofolate reductase as a standard while passing through the solution gel filtration chromatography.

公开(公告)号：JPH0698766A

公开(公告)日：1994-04-12

申请号：JP15557591

申请日：1991-05-30

Applicant: AGENCY IND SCIENCE TECHN

Inventor： HIRANO TAKASHI ,  TODOROKI TAKESHI ,  OHASHI SHINICHI

IPC: A61K38/44 ,  A61P29/00 ,  C12N9/02

Abstract: PURPOSE:To obtain a polymer derivative having high activity and excellent anti-inflammatory action by adding a dimethylmaleic acid anhydride solution to a superoxide dismutase solution, reacting the components and bonding the reaction product to a divinylethyl ether-maleic anhydride copolymer, etc. CONSTITUTION:A superoxide dismutase(SOD) is dissolved in a weakly basic inorganic buffer solution, an organic solvent solution of dimethylmaleic acid anhydride of formula I is dropped to the above solution and made to react with each other, the reaction product is incorporated and bonded with an organic solvent solution of a polymer such as a divinylethyl ether-maleic anhydride copolymer (DIVEMA) expressed by formula II (m is integer of 20-500) or a polyethylene glycol derivative (PEG2) expressed by formula III (n, is integer of 10-700) and the reaction solution is adjusted to weakly acidic state. The precipitated dimethylmaleic acid is separated and the reaction product is purified to obtain an SOD-polymer derivative expressed by formula IV [Polymer is group of formula V (m is integer of 20-500; the group is bonded to the amino group of SOD at the position of X through amide bond); (i) is integer of 1-22].

公开(公告)号：JPH0698762A

公开(公告)日：1994-04-12

申请号：JP40100490

申请日：1990-12-10

Applicant: HITACHI CHEMICAL CO LTD ,  AGENCY IND SCIENCE TECHN

Inventor： TAKASUKA AKIKO ,  IZUTSU HIROSHI ,  YOSHINO KATSUHIKO ,  IWAKURA MASAHIRO ,  OHASHI SHINICHI ,  TANAKA YOSHIO

IPC: A61K38/00 ,  A61P37/08 ,  C07K1/22 ,  C07K14/00 ,  C07K19/00 ,  C12N1/21 ,  C12N15/09 ,  C12N15/62 ,  C12P21/02 ,  C12R1/19 ,  A61K37/02 ,  C07K13/00

Abstract: PURPOSE:To produce a large amount of DSDGK by expressing a macromer of an antiallergic pentapeptide having an amino acid sequence of Asp-Ser-Asp- Gly-Lys (DSDGK by single letter denotation) as a fused protein with dihydrofolic acid reductase (DHFR). CONSTITUTION:A single-stranded DNA composed of two or more recurring units consisting of a base sequence (expressed by P and Q) coding DSDGK is mixed with a single-stranded DNA complemental to the former DNA. The mixture is cooled, four kinds of bases are made to react in the presence of a DNA polymerase, the reaction system is heated and optionally a part of the above reaction is repeated to obtain a DNA chain having a large amount of base sequences coding DSDGK on the same chain in repeated manner. The DNA chain is inserted into a vector containing a base sequence coding DHFR to construct a new recombinant vector capable of expressing a fused protein (IV) of a DHFR-DSDGK macromer.

公开(公告)号：JPH04187091A

公开(公告)日：1992-07-03

申请号：JP31594990

申请日：1990-11-22

Applicant: HITACHI CHEMICAL CO LTD ,  AGENCY IND SCIENCE TECHN

Inventor： TAKASUKA AKIKO ,  IZUTSU HIROSHI ,  YOSHINO KATSUHIKO ,  IWAKURA MASAHIRO ,  OHASHI SHINICHI ,  TANAKA YOSHIO

IPC: C12P21/02 ,  C07K1/22 ,  C07K7/06 ,  C07K14/00 ,  C07K14/195 ,  C07K14/41 ,  C07K19/00 ,  C12N9/04 ,  C12N15/09 ,  C12N15/53 ,  C12N15/62 ,  C12R1/19

Abstract: NEW MATERIAL:A recombinant vector containing a base sequence encoding a fused protein with a polymer of dihydrofolate reductase-antiallergic pentapeptide polymer. EXAMPLE:Recombinant plasmid pD314TX. USE:For producing DSDGK. PREPARATION:A recombinant plasmid shown by the example is obtained by the scheme shown by the formula.

公开(公告)号：JPH04117285A

公开(公告)日：1992-04-17

申请号：JP12320290

申请日：1990-05-15

Applicant: AGENCY IND SCIENCE TECHN ,  HITACHI CHEMICAL CO LTD

Inventor： BABA KENZO ,  IWAKURA MASAHIRO ,  OHASHI SHINICHI ,  TANAKA YOSHIO ,  IZUTSU HIROSHI ,  OBARA KAZUHIKO ,  TAKASUKA AKIKO

IPC: C12N15/09 ,  A61K38/00 ,  A61K38/44 ,  A61P37/08 ,  C07K1/12 ,  C07K1/22 ,  C07K7/06 ,  C07K14/00 ,  C07K19/00 ,  C12N1/21 ,  C12N9/02 ,  C12N15/12 ,  C12N15/53 ,  C12N15/62 ,  C12P21/02 ,  C12R1/19

Abstract: NEW MATERIAL:A recombinant plasmid containing a base sequence encoding dihydrofolate reductase-antiallergic peptide polymer fused protein (I). EXAMPLE:A recombinant plasmid wherein an antiallergic pentapeptide, a fused protein (I) of dimer contains an amino acid sequence shown by the formula (underline shows amino acid existing between dihydrofolate reductase and antiallergic pentapeptide). USE:Production of DSDGK. PREPARATION:A DNA encoding DSDGK polymer obtained by chemically synthesizing a single strand DNA and annealing both chains is inserted into a vector plasmid capable of manifesting DHFR.

公开(公告)号：JPH0279978A

公开(公告)日：1990-03-20

申请号：JP23124888

申请日：1988-09-14

Applicant: AGENCY IND SCIENCE TECHN

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

IPC: C12N15/09 ,  A61K38/22 ,  A61P23/00 ,  A61P25/04 ,  C07K1/16 ,  C07K1/22 ,  C07K14/00 ,  C07K14/575 ,  C07K14/655 ,  C07K14/675 ,  C07K19/00 ,  C12N1/20 ,  C12N1/21 ,  C12N15/16 ,  C12N15/62 ,  C12P21/02 ,  C12R1/19

Abstract: PURPOSE:To obtain a recombinant plasmid pENDD2 being a recombinant plasmid having 4682 pair of bases and capable of producing delta-endorphin of morphine like peptide having high analgesic activity when Escherichia coli is transformed by the above-mentioned plasmid. CONSTITUTION:Gene coding delta-endorphin expressed by the formula is synthesized and the synthesized gene is integrated into a plasmid pMEK2 to prepare a fused gene (pENDCD2) of delta-endorphin gene and dihydrofolic acid reducing enzyme (DHFR) gene, by which Escherichia coli is then transformed. DHFR-delta- endorphin fused protein is produced by culturing the transformed Escherichia coli and then the fused protein is digested by a proteolytic enzyme to separate delta-endorphin.

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.

公开(公告)号：JPH01144995A

公开(公告)日：1989-06-07

申请号：JP30215587

申请日：1987-11-30

Applicant: AGENCY IND SCIENCE TECHN

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

IPC: C07K14/655 ,  C07K1/16 ,  C07K1/22 ,  C07K14/00 ,  C07K14/195 ,  C07K14/41 ,  C07K14/575 ,  C12N9/06 ,  C12N15/09 ,  C12P21/02 ,  C12R1/19

Abstract: PURPOSE:To enable the separation and purification of a dihydrofolic acid reductase-somatostatin fused protein produced by E.coli transformed with plasmid pTPGIF2, by purifying the protein using a specific chromatography. CONSTITUTION:Escherichia coli containing plasmid pTPGIF2 (FERM BP-1577) is cultured and the obtained bacterial cells are disintegrated and centrifuged. The cell-free extraction liquid of the supernatant is subjected to ion-exchange column, methotrexate-bonded affinity column chromatography and anion- exchange column chromatography using dihydrofolic acid reductase activity as a criterion. The objective dihydrofolic acid reductase-somatostatin fused protein shown in the figure can be produced by this process in purified state.

公开(公告)号：JPH01144978A

公开(公告)日：1989-06-07

申请号：JP30215687

申请日：1987-11-30

Applicant: AGENCY IND SCIENCE TECHN

Inventor： IWAKURA MASAHIRO ,  KOKUBU TOMOKUNI ,  FURUSAWA KIYOTAKA ,  OHASHI SHINICHI ,  SAKAI TSUKASA ,  TANAKA YOSHIO

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

Abstract: PURPOSE:To produce a peptide (GRF1-29) containing the factors from the 1st to the 29th of bovine growth hormone releasing factors, by using a specific recombinant plasmid pGRE2-15. CONSTITUTION:A recombinant plasmid is prepared by linking a GRF1-29 gene to the 3'-terminal of a dihydrofolic acid reductase (DHFR) gene using a plasmid vector pTP70-1 capable of fusing a heteropeptide to a carboxy-terminal or DHFR of E.coli. The plasmid is introduced into E.coli and expressed to enable the stable production and accumulation of a fused protein containing GRF1-29 bonded to the carboxy-terminal of DHFR in E.coli.

公开(公告)号：JPS63223084A

公开(公告)日：1988-09-16

申请号：JP5615487

申请日：1987-03-11

Applicant: AGENCY IND SCIENCE TECHN

Inventor： TAMAOKI TAKASHI ,  OHASHI SHINICHI ,  SAKURAGI MASAKO ,  ICHIMURA KUNIHIRO ,  ARIMA IKUKO

IPC: C09K9/02 ,  G03C1/685

Abstract: PURPOSE:To obtain a photochromic composition having excellent light durability, coloring by ultraviolet light, decoloring by irradiation with visible light ray, by including a specific spirobenzopyran compound in a specific (modified) gamma- cyclodextrin. CONSTITUTION:A spirobenzopyran compound shown by formula I (R1 is H, halogen, methyl or methoxy; R2 is nitro or halogen) is included in (B) a (modified) gamma-cyclodextrin shown by formula II (R1 is H, methyl or naphthyl group- containing residue; R2 and R3 are H or methyl; n is 8) to give the aimed photochromic composition.