| Project/Area Number |
63490004
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
広領域
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| Research Institution | University of Tsukuba |
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Principal Investigator |
HARADA Hiroshi University of Tsukuba, Institute of Biological Sciences, Professor, 生物科学系, 教授 (90015991)
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| Co-Investigator(Kenkyū-buntansha) |
KAMADA Hiroshi University of Tsukuba, Gene Experiment Center, Associate Professor, 遺伝子実験センター, 助教授 (00169608)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1988: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | Pollen meiosis / pollen mother cell protein (PMCP) / Hl histone / nucleosome / (PMCP) / Hlヒストン / HIヒストン / ポリクローナル抗体 / ヌクレオゾーム |
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| Research Abstract |
In microsporogenesis of lily PMCP(Pollen Mother Cell Protein) was detected after preneiosis stage. We analyzed PMCP localization on nucleosone Nucleosome sonomer,diner and trimer could be isolated with 4-25% linear sucrose gradient for 24hr centrifugation (25,000 rps) at 4ﾟC. Histones were observed in all fractions. PMCP was detected In nucleosome monomer by SDS-PAGE analysis. It is possible that monomer containing PMCP delayed on native 5% polyscrylaside gel electrophoresis. Because nobility of nucleosome depends on their molecular size,charge and so on.
We measured PMCP content per nucleosome monomer,diner and trimer with densitometer.
The PMCP contents of diner and trimer are two and three fold higher than that of monomer, respectively, as based on the contents of the H4 histone. PMCP contents of polymers more than trimer are nearly comparable to that of trimer in this experiment. From densitometrical analysis, it is suggested that PMCP localizes on nuoleosome linker domain. PMCP may play an important role for organization of high order structure of chromatin with another mode than HI histone.
Finally, we tried to determine partial amino acid sequence of PMCP. PMCP and Hl were isolated by HPLC and applied to Protein sequense . However,amino acid signal could not be appeared. This failure is due to the blockins of N terminal amino acid. Then we examined cleavage of PMCP. Chemical reagent such as acetic acid, N-Bromosuccinicimidre, and cyanogen bromide were useful for cleavage of PMCP to peptide .
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