| Project/Area Number |
01480539
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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 |
分子遺伝学・分子生理学
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| Research Institution | Osaka University |
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Principal Investigator |
OGAWA Tomoko Osaka Univ., Dept. of Bio., Asistant Professor, 理学部, 講師 (80028208)
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| Co-Investigator(Kenkyū-buntansha) |
OGAWA Hideyuki Osaka Univ., Dept. of Biol., Professor., 理学部, 教授 (70028207)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1991: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1990: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1989: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Keywords | RecA Protein / RecA Protein Structure / Chimeric gene. / Chimeric RecA Protein / RecBCsbcA gene in E. coli. / Recombination / Oligomerization of RecA Protein / RecA gene of Pseudomonas aeruginosa / SOSresponce / recAキメラ遺伝子 / recBrecCsbcA変異株 / 遺伝子組換え / SOS機能の誘導 / RecA蛋白質の機能ドメイン / キメラ遺伝子形成の新方法 / 大腸菌recA遺伝子 / 緑膿菌recA遺伝子 / 大腸菌・緑膿菌のキメラrecA遺伝子 / ドメイン構造 / RecA蛋白質のトリプシン部分分解 / DNA結合部位 / ATP結合部位 / 一本鎖DNA依存ATPase |
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| Research Abstract |
We developed a novel genetic method for finding functional regions of a protein by the analysis of chimeras formed between homologous proteins. Sets of chimeric genes were made by intramolecular homologous recombination in a linearized plasmid DNA carrying both recA genes of Escherichia coli and Pseudomonas aeruginosa. A recBCsbcA strain of E. coli was used for isolation of plasmids carrying recombinants between these genes. Examination of properties of E. coli strains deleting the recA gene and carrying a plasmid with a chimeric gene shows that chimera formation at certain positions inactivates a RecA function. Frequently, all chimeras with a junction in a certain region of the protein inactivate a function. Rather than a direct effect of the presence of the junction at a particular position, mismatching of the regions both sides of the junction that are derived from the different species is responsible for the inactivation. For a chimeric protein to be functional, certain pairs of sequences in different regions of the protein must derive from the same parent. Four pairs of such sequences were found : two are involved in activities for genetic recombination and for resistance to ultraviolet light irradiation and the others in formation of active oligomera. Regions defined by these sequences are located in the looped regions of the protein. A pair of regions may cooperate to form a functional folded structure.
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