| Project/Area Number |
61570206
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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 |
細菌学
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| Research Institution | Osaka University |
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Principal Investigator |
IMAGAWA Tadashi (Medical School, Research Associate) Osaka University, 医学部, 助手 (10036478)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Bordetella pertussis / Adenylate Cyclase / Gene / クローニング / 遺伝子のクローニング |
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| Research Abstract |
The gene library of chronosomal DNA from Bordetella pertussis strain Tohama was constructed using pBR322 vector in E.coli cya mutant derived from strain x1776 in this project and screened for the adenylate cyclase gene to confer Cya^+ phenotype to the mutant. The hybrid plasmid which complements the phenotype of the cya mutant harbored an insert DNA fragment with a molecular size of 3.7 Kb. This hybrid plasmid confer Cya^+ phenotype also to another <delta>cya mutant strain TP2010 of E.coli derived from Dr.A.Danchin. Analysis by Southern hybridization revealed that the 3.7 Kb insert derived from B.pertussis DNA was weekly but significantly hybridised with ^<32>Plabeled 6 Kb insert in pCA2 including E.coli adenylate cyclase gene as probe which was obtained from Dr.H.Aiba (Tsukuba Univ.), indicating that there is some significant homology between two species of adenylate cyclase gene. Nevertheless,reproducible results ofn asssay for adenylate cyclase activity have never been obtained in E.coli <delta>cya mutant harboring 3.7 Kb fragment inserted into several kinds of expression vector. Nucleotide sequence amalysis is now in progress for 3.7 Kb insert DNA using dideoxy method.
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