| Project/Area Number |
11670145
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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 |
Pathological medical chemistry
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| Research Institution | Saga Medical School |
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Principal Investigator |
MUKAI Tsunehiro Saga Medicl School, Department of Biochemistry, Professor, 医学部, 教授 (40108741)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | genomic imprinting / p57Kip2 mutant / transgenic mouse / Beckwith-Wiedemann syndrome / インプリンティング / p57KIP2変異体 / ゲノムインプリンティング / P57KIP2 / アデノウイルスベクター |
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| Research Abstract |
The CDK inhibitor, p57^<KIP2> is one of the gene responsible for Beckwitn-Wiedemann syndrome (BWS) of which locus is located on chromosome 11p15.5. To confirm the phenotype of this mutant at the cellular level and the animal, 1) we transfected the mutant to the cell and observed the biochemical and morphorogical effect, 2) and also created the transgenic mouse as a model of BWS.Adenovirus vector carrying mutant p57^<KIP2> cDNA was transfected to the G401 and RD cells. These cells do not express the endogenous p57KIP2. The mutant is Ad/KIP2-1-8 (mutaion at the C-ter of p57^<KIP2>). Controls are Ad/KIP2-1 (wild p57^<KIP2>) and Ad/LacZ.RD cell transfected with wild p57^<KIP2> cDNA expressed the protein. This was confirmed by Western blotting and by the staining with p57KIP2 antibody. However, the specific repression of the cell growth was not observed by the wild p57^<KIP2>. The level of the repression was similar with that of the mutant p57^<KIP2>. Next, we tried to create the transgenic mouse carrying mutant p57^<KIP2> as animal model to mimic BWS.pRc/CVM plasmid vector was used to obtain the high expression of human cDNA recombinant. These include pDR2-1 (wild), pDR2-1-6 (mutation of the CDK inhibitory domain) and pDR2-1-8 (mutaion at the QT domain). The obtained result showed all mice born were non-transgenic, suggesting the overexpression of this gene might be toxic. Conditional transgenics such as Cre/Lox system will be necessary to overcome this situation in the future.
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