1987 Fiscal Year Final Research Report Summary
Bacterial differentiation: Its application on production of useful substances.
| Project/Area Number |
60303024
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| Research Category |
Grant-in-Aid for Co-operative Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
発酵・醸造
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
KOBAYASHI Yasuo Hiroshima Univ., Fac. Appl. Biol. Sci., 生物生産学部, 教授 (10013319)
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| Co-Investigator(Kenkyū-buntansha) |
SADAIE Yoshito Natl. Inst. Genet., 助教授 (40000252)
YAMANE Kunio Univ. of Tsukuba, Inst. Biol. Sci., 生物科学系, 助教授 (20013336)
YAMASAKI Makari Univ. of Tokyo, Fac. Agr., 農学部, 教授 (60011889)
BEPPU Teruhiko Univ. of Tokyo, Fac. Agr., 農学部, 教授 (80011873)
SAITO Hiuga Univ. of Tokyo, Inst. Appl. Microiol., 応用微生物研究所, 教授 (10013301)
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| Project Period (FY) |
1985 – 1987
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| Keywords | Bacillus subtilis / sporulation / catabolite repression / A-factor / promoter-probe vector / secretion vector / gene amplification / プテアーゼ産生 |
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| Research Abstract |
1. Control mechanism of bacterial differentiation (Saito, Sadafe, Fujita, Ikeuchi, Kawamura) (1) Regulatory mechanism of sporulation initiation gene spoOA expression in Bacillus subtilis was studied by the use of lacZ-fusion gene and temperature-sensitive spoOA mutant. It was shown that expression of spoOA gene depends on spoOA, OB, OE, OF, and OH genes and spprulation was not initated unless the function of SpoOA protein is derepressed. (2) spoOA gene was cloned and its gene product was overproduced in Escherichia coli and purified. Purified SpoOA protein has DNA-binding activity. (3)div gene which regulates cell division, sporulation, and protein secretion was cloned. (4) Catabolite-repressible gluconate operon was cloned. The whole nucleotide sequence was determined and the mechanism of catabolite repression of B. subtilis was studied.
2. Isolation of regulatory factor of gene expression (Beppu, Nakayama) (1) Streptomycin production and sporulation in Streptomyces griseus is positively controlled by A-factor. A-factor was purified and regulatory network surrounding A-factor was studied. (2) Chromosomal proteins of B. subtilis were isolated and purified and sporulation-specific chromosomal proteins were identified.
3. Development of usefulvector (Kobayashi, Yamasaki, Yamane, Kudoh, Tanaka) (1) Isolation and analysis of a temperature-dependent promoter revealed that its expressio is controlled by translational-coupling. Stable promoter-probe vector for B. subtilis was constructed. (2) A commom method to amplify a useful gene in B. Subtilis chromosome was developed. (3) Secretion vector carrying -amylase signal peptide was constructed and improvement of its secretion ability was intended. (4) prtR gene which increases protease production in B. natto was identified and cloned. It was shown that PrtR protein is a positive factor which stimulates the transcription of protease gene.
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