| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Research Abstract |
Dr.Mayr found that there were similar about 100 amino acid sequences in various signaling molecules, which have divergent function. He called this domain as pleckstrin homology (PH) one, and proposed that this domain wouldbe important motif on "Cell " journal at 1993. When I planned this project, only 6 proteins which contained PH domain were reported. However, since researchers recognized PH domain as valuable, the number of PH containing protein rapidly increased.
My plan was 1) molecular cloning of chicken pleckstrin cDNA,2) molecular cloning of genomic pleckstrin DNA,3) gene knockout in chicken B lymphocyte DT40 and functional analysis. At first, I tried to molecular cloning of chicken pleckstrin cDNA.In consequence of several efforts, I took several cDNA clones and analyzed them. Positive clone contained an 1,059 bp open reading frame encoding a polypeptide of 353 residues sharing 81% overall identity with the human pleckstrin. Then N- and C-terminal PH domain was also conserved ; 89% and 79%, respectively. The length of chicken pleckstrin's mRNA was about 3kb and this expression was restricted to spleen and DT40 B cell line. Then, using this cDNA as a probe, I took several genomic clones. But they are too complicated to analyze, I could not completely finish this No.2) project. Therefore I could not undertake the plan 3) gene knockout in chicken B lymphocyte DT40 and functional analysis.
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