| Project/Area Number |
10672077
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | Kobe Gakuin University |
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Principal Investigator |
OKAMOTO Hiroshi Kobe Gakuin University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (00028870)
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| Co-Investigator(Kenkyū-buntansha) |
YAYAMA Katsutoshi Kobe Gakuin University, Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助手 (30248108)
TAKANO Masaoki Kobe Gakuin University, Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助手 (30258107)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1998: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | kininogen / Kinin / Kallikrein-Kinin System / ブラジキニン / カリクレイン・キニン系 / ノックアウトマウス |
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| Research Abstract |
Mammalian expresses two types of protein precursor for kinins termed high-molecular-weight and low-molecular-weight kininogens which are coded by a single gene and produced by alternative splicing of the gene transcript. To clarify roles of the kallikrein-kinin system in the physiological and pathological conditions, steps have been taken toward the generation of mice with a "knockout" of the kininogen gene. A genomic clones of the mouse kininogen were isolated from129 strain mice using LA-PCR method and their coding sequences determed by DNA sequence analysis. A physical map of the DNA flanking this coding sequence was generated. According to the structure of kininogen gene, a vector was constructed for targeted disruption of the mouse kininogen gene in plasmid pPNT. This vector contains 7.5 Kb of DNA upstream from the bradykinin coding sequence, a neomycin resistance gene, and 2.4 Kb of DNA downstream from bradykinin coding sequence. Thus, the correct homologous recombination event will result in a chromosome in which the coding sequence for the bradykinin in mouse kininogen is replaced with the neomycin resistance gene. This targetting vector has been transfected into embryonic stem cells, and clones containing a targeted disruption of the mouse kininogen will be identified.
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