| Project/Area Number |
62470117
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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 |
応用生物化学・栄養化学
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
MIZUNO Shigeki Professor, Faculty of Agriculture, Tohoku University, 農学部, 教授 (90112903)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIHARA Hiroaki Assistant, Faculty of Agriculture, Tohoku University, 農学部, 助手 (50005633)
吉田 充輝 東北大学, 農学部, 助教授 (20005648)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1988: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | fibroin H- and L-chain genes / sequence-specific DNA binding protein / middle and posterior silk glands of silkworm / bent-repetitive DNA / W chromosome / W-protein / 反復塩基配列 / W染色体 / W-protein / クロマチン / フィブロインL鎖、H鎖遺伝子 / カイコ / 中部、後部絹系腺 / ニワトリ / 湾曲反復DNA / 非ヒストンタンパク質 / Wーprotein / DNaseIフットプリント法 / ゲルシフト法 / 遺伝子の同調的発現制御 / 反復DNA配列 |
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| Research Abstract |
1. Protein factors binding to the 5'-upstream region of the fibroin light(L-) chain gene: Sequence analysis of the cosmid clone containing the fibroin L-chain gene revealed that three sequences in the 5'-upstream region, i.e, -415- -398, -306- -277, -250- -226, were highly homologous to the sequences in the -275- -155 region of the fibroin H-chain gene. It was found that protein factors which bound to the all three upstream regions were present in the 0.5M NaCl extract of the posterior silk gland nuclei of the silkworm, Bombyx mori. Particularly, proteins binding to the 64bp sequence containing the -306- -277 sequence were shown to consist of posterior silk gland-specific factor(s) and posterior and middle silk gland-common factor(s).
2. Purification and properties of W-protein: W-protein, purified from the 0.35M NaCl extract of the chicken liver nuclei, has a monomeric molecular weight of 72k but forms a multimeric form consisting of about 30 molecules, under non-denaturing conditions. W-protein binds with high affinity to the chicken W chromosome-specific, bent repetitive DNA(XhoI family). It was suggested that the XhoI family sequence wraps around the multimer of W-protein making multiple contacts through the A-T-rich minor grooves..participation of W-protein in the formation of higher-order structure of chromatin is suggested.
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