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    NOVEL RECOMBINANT PLASMID PBSFOLEK1
    1.
    发明专利
    NOVEL RECOMBINANT PLASMID PBSFOLEK1 失效

    公开(公告)号：JPS6387981A

    公开(公告)日：1988-04-19

    申请号：JP23400386

    申请日：1986-09-30

    Applicant: AGENCY IND SCIENCE TECHN

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

    IPC: C12N15/09 ,  C07K14/665 ,  C12N1/20 ,  C12N1/21 ,  C12N9/06 ,  C12N15/00 ,  C12P21/02 ,  C12R1/19

    Abstract: PURPOSE:To provide a recombinant plasmid containing integrated gene of dihydrofolic acid reductase (DHFR)-leucine encephaline fused protein, stably replicable in E.coli and capable of imparting said host with trimethoprim resistance and ampicillin resistance. CONSTITUTION:Plasmid pBSDHFR1 is cut with restriction enzymes EcoRI and PstI and separated by 1% agarose gel electrophoresis to obtain a small fragment DNA. Separately, plasmid pBR322 is cut with restriction enzyme BamHI and Pst and separated by 1% agarose gel electrophoresis to obtain a large fragment DNA. The DNA of formula I and formula II are synthesized, the 5'-terminal is converted to phosphoric acid group and the product is annealed to obtain a DNA coding leucine encephaline. The above 3 kinds of DNA molecules are linked together, introduced into E.coli C600 strain and cultured. The objective recombinant DNA containing the sequence of formula III coding a DHFR-leucine encephaline fused protein can be separated from the obtained colony.

    2'-O-TETRAHYDROPYRANYLNUCLEOSIDE DERIVATIVE AND ITS PREPARATION
    2.
    发明专利
    2'-O-TETRAHYDROPYRANYLNUCLEOSIDE DERIVATIVE AND ITS PREPARATION 失效

    公开(公告)号：JPH01224391A

    公开(公告)日：1989-09-07

    申请号：JP4927388

    申请日：1988-03-02

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： FURUSAWA KIYOTAKA ,  SAKAI TSUKASA

    IPC: C07H19/067 ,  C07H19/167 ,  C07H23/00

    Abstract: NEW MATERIAL:A compound of formula I [B is nucleic acid base which may be protected; R, R' are phenyl, branched alkyl or 3 or more carbon atoms.; R'' is tetrahydropyranyl residue]. EXAMPLE:2'-O-tetrahydropyranyl-3',5'-O-(di-tert.-butylsilandiyl)-uridine. USE:An intermediate of RNA-relating compounds in genetic engineering. PREPARATION:The nucleoside of formula II is silylated with a difunctional silicon compound of the formula: RR'SiX2 (X is acid residue), then allowed to react with dihydropyran.

    MANIFESTATION VECTOR PTP104-4
    3.
    发明专利
    MANIFESTATION VECTOR PTP104-4 失效

    公开(公告)号：JPH01144979A

    公开(公告)日：1989-06-07

    申请号：JP30215787

    申请日：1987-11-30

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： IWAKURA MASAHIRO ,  FURUSAWA KIYOTAKA ,  SAKAI TSUKASA ,  TANAKA YOSHIO

    IPC: C12N1/21 ,  C12N15/00 ,  C12N15/09 ,  C12N15/53 ,  C12N15/62 ,  C12P21/00 ,  C12P21/02 ,  C12R1/19

    Abstract: PURPOSE:To produce a substance containing a heterogenic product fused to a carboxy-terminal of dihydrofolic acid reductase(DHFR), in high efficiency, by modifying a plasmid pTP70-1 capable of easily utilizing a DHFR gene as a genetic marker. CONSTITUTION:A terminator region of rrnB gene (one of ribosomal RNA genes) known as an efficient terminator is introduced at the downstream side of a DHFR gene of plasmid pTP70-1 to obtain a plasmid pTP104-4 capable of efficiently producing a substance containing heterogenic product fused to the carboxy-terminal of DHFR. The plasmid has a size of 4,466 base pairs and can impart a host E.coli with trimethoprim resistance and ampicillin resistance.

    PRODUCTION OF O-SILYLATED HYDROXY-CONTAINING CYCLIC ETHER
    5.
    发明专利
    PRODUCTION OF O-SILYLATED HYDROXY-CONTAINING CYCLIC ETHER 失效

    公开(公告)号：JP2000053694A

    公开(公告)日：2000-02-22

    申请号：JP22497998

    申请日：1998-08-07

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： FURUSAWA KIYOTAKA

    IPC: C07H23/00 ,  C07H11/00 ,  C07H19/06 ,  C07H19/16

    Abstract: PROBLEM TO BE SOLVED: To easily produce an O-silylated hydroxy-containing cyclic ether useful as a raw material for pharmaceuticals, etc., in one-pot at a low cost dispensing with troublesome separation procedure by successively reacting a specific OH- containing cyclic ether with a bifunctional and monofunctional silicon compounds and reacting the product with hydrofluoric acid. SOLUTION: The objective compound of formula III (when Y1 or Y2 is hydroxymethylene, the OH group is converted to OSiR4R5R6) is produced by reacting (A) a compound of formula R2R3SiX12 (R2 and R3 are each an aromatic group or a >=3C branched aliphatic group; X1 is a residue of an acid other than hydrochloric acid) with (B) a compound of formula I (R1 is a substituent free from reactivity with the component A; Y1 and Y2 are each methylene, hydroxymethylene or R1-substituted methylene; (t) and (q) are each 0 or 1) e.g. a compound of formula II [B is a (protected) nucleic acid base]}, reacting with (C) a compound of formula R4R5R6SiX2 (R4 to R6 are each an aromatic group or an aliphatic group; X2 is an eliminable group) and finally reacting with (D) hydrofluoric acid in the presence of (E) a base.

    DEPROTECTION OF CYCLIC SILICON PROTECTING GROUP
    6.
    发明专利
    DEPROTECTION OF CYCLIC SILICON PROTECTING GROUP 失效

    公开(公告)号：JPH01230595A

    公开(公告)日：1989-09-14

    申请号：JP5717388

    申请日：1988-03-10

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： FURUSAWA KIYOTAKA ,  SAKAI TSUKASA

    IPC: C07H19/06 ,  C07H19/16 ,  C07H23/00

    Abstract: PURPOSE:To eliminate the title silicon group, by hydrolyzing a specific cyclic silicon derivative to cut several sites among siloxane bonds preferentially whereby local eliminating can be done highly selectively, thus the technique can be beneficially applied to biotechnology. CONSTITUTION:The cyclic silicone derivative of the formula (B is unprotected or protected nucleic acid base; R, R' are phenyl, branched alkyl of 3 or more carbon atoms; R'' is H, unprotected or protected hydroxyl) is hydrolyzed with an acid such as hydrochloric acid or an alkali such as sodium hydroxide to cut one bond in 2 siloxane bonds preferentially whereby the objective silicon group is eliminated.

    2'-O-SILYL CYCLIC SILICATED NUCLEOTIDE DERIVATIVE, PRODUCTION THEREOF AND PRODUCTION OF 2'-O-SILYL NUCLEOTIDE THEREFROM
    10.
    发明专利
    2'-O-SILYL CYCLIC SILICATED NUCLEOTIDE DERIVATIVE, PRODUCTION THEREOF AND PRODUCTION OF 2'-O-SILYL NUCLEOTIDE THEREFROM 失效

    公开(公告)号：JPH10226697A

    公开(公告)日：1998-08-25

    申请号：JP3380897

    申请日：1997-02-18

    Applicant: AGENCY IND SCIENCE TECHN

    Inventor： FURUSAWA KIYOTAKA

    IPC: C07H19/067 ,  C07H19/167 ,  C07H23/00

    Abstract: PROBLEM TO BE SOLVED: To obtain the subject new derivative useful as an intermediate for synthesizing and modifying ribonucleic acid compounds, etc., by cyclizing a nucleotide with a bifunctional silicon compound and subsequently silylating the 2-hydroxyl group of the cyclized silyl compound with a monofunctional silicon compound. SOLUTION: A new 2'-O-silyl cyclic-silicified nucleotide derivative represented by formula I [B is a (protected) nucleic acid base; R , R are each an aromatic group, a >=3C branched aliphatic group; R to R are each an aromatic group, an aliphatic group] [e.g. 2'-O-tert-butyldimethylsilyl-3',5'-O-(di-tert-butylsilanediyl) uridine]. The compound of formula I is useful as an intermediate for synthesizing and modifying ribonucleic acid compounds, etc. The compound of formula I is obtained by silylating a nucleotide of formula II with a bifunctional silicon compound of the formula: R R SiX 2 (X is an acid residue) and further silylating the remaining 2'-hydroxyl group of the obtained cyclic compound with a monofunctional silicon compound of the formula: R R R SiX (X is a releasable group).

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