| Project/Area Number |
08671162
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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 |
内分泌・代謝学
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| Research Institution | KOBE UNIVERSITY |
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Principal Investigator |
ABE Hiromi KOBE UNIVERSITY FACULTY OF MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (20159437)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1997: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | Somatostatin / Somatostatin receptors / Knockout mouse / GHRH / Growth Hormone / Pituitary / Hypothalamus / 2型ソマトスタチン受容体 / 5型ソマトスタチン受容体 / ジーンターゲッテング / 視床下部ペプチド |
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| Research Abstract |
We subcloned genomic DNA fragment of somatostatin receptor, SSTR2 and SSTR5, with PCR using several sets of oligoprimers. 3.8Kb Fragment of SSTR2 and 4.2Kb fragment of SSTR 5 were obtained. Both fragment consist with small portion of 5'franking sequence and receptor coding regions. Neo-cassette was inserted into the coding region and the fragments were subcloned into the pMCtk vector with HSV-tK.Those vectors were transfected to the ES cells, then, clones were selected by Neo-resistance first followed by positive selection with acyclovir. After this step, 35 clones were selected and PCR with primers outside the homologously recombined site revealed that 3 out of 35 clones were successfully recombinated with trasfected DNA Fragment (two clones for SSTR2 and one for SSTR5). These 3 clones were injected to mouse embryo. We got heterozygous F0 judging from the fur patches, however, failed to get homozygous knockout mice in F1 generations.
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