| Project/Area Number |
04640620
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物生理学
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| Research Institution | Tokyo Gakugei University |
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Principal Investigator |
SATO Naoki Tokyo Gakugei University, Ass.Prof., 教育学部, 助教授 (40154075)
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| Co-Investigator(Kenkyū-buntansha) |
KUROIWA Tsuneyoshi University of Tokyo, Fac.of Science, Prof., 大学院・理学系研究科, 教授 (50033353)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | chloroplast / envelope / DNA-binding protein / nucleoid / エンドウ葉緑体 / 葉緑体包膜 / サウスウェスタン法 / サイトカイニン / 葉緑体DNA / PENDタンパク質 |
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| Research Abstract |
1.Dynamics of cpDNA.Observation by fluorescence microscopy of the binding of cpDNA with plastid envelope at various stages of development of etioplast and chloroplast indicated that cpDNA binds to plastid envelope in young plastids active in division and that this binding was not affected by light or the presence of thylakoid membrane.
2.Identification of a cpDNA-specific DNA-binding protein in plastid envelope. A 130 kDa DNA-binding protein was detected in plastid envelop by South-western technique. The protein named PEND protein was localized in the inner envelope, and was able to bind to several (at least four) distinct regions of cpDNA.PEND protein was detected in the envelope of young plastid but not in the envelop of mature chloroplast.
3.Effort to purify PEND protein. PEND protein was partially affinty-purified using a biotin-labelled DNA probe and streptavidin-coated magnetic particles. The protein was detected as a 115kDa polypeptide in the standard SDS-PAGE.
4.cDNA cloning of plastid DNA-binding proteins. A series of South-western screening yielded 3 clones of cDNA.The antibody against one of these proteins (PD1) detected 50kDa and 87kDa polypeptides in the plastid. The accumulation of these polypeptides was markedly reduced by a cytokinin.
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