| Project/Area Number |
06670314
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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 |
Bacteriology (including Mycology)
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| Research Institution | Obihiro University of Agricuruture and Veterinary Medicine (1995)
国立公衆衛生院 (1994) |
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Principal Investigator |
MAKINO Sou-ichi Obihiro University of Agricuruture and Veterinary Medicine, Department of Veterinary Medicine, Assistant Professor, 畜産学部, 助教授 (30181621)
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| Co-Investigator(Kenkyū-buntansha) |
OKADA Yumiko The Institute of Public Health, Department of Veterinary Public Health, Research, 衛生獣医学部, 研究官 (50232137)
SASAKAWA Chihiro Institute of Medical Science, University of Tokyo, Professor, 医科学研究所, 教授 (70114494)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1994: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Listeri monocytogenes / Virulence / NaCl / PCR / flagella / detection / 低温増殖能 / 鞭毛 / 病原性 / 食品 |
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| Research Abstract |
Molecular genetical analysis of the growth at low temperature and at high concentration of NaCl in Listeria monocytogenes have been studied. We focused on the formation of flagella at low temperature. Similtaneously, we have established the direct detection system of L.monocytogenes DNA from food by PCR.
1.Anti-fkagella antisurum : Using flagella protein isolated from SDS-polyacrylamidgel elecrtophoresis, We have made anti-rabbit serum raised against flagellin (anti-fla).
2.Cloning of flagella-related genes by anti-fla : In E.coli, we have isolated a several clones positively reacted on the anti-fla, followed by the genetical analysis and immunological analysis. Although one clone was the structural gene for flagella-formation, other 2 clones were new genes different from the flagella gene. But the both clones, clone 1 and clone 2, produced their gene products reacted against anti-fla by western blotting analysis.
3.fla-minus mutant : We constructed fla-minus mutant, followed by invasion tests in the epithelial cells and infection tests to mouse. But, we could not detect any defference between the mutant and its parent strain.
4.PCR method : Using the nucleotide sequence of clone 1, we constructed a set of primer for Listeria species. By PCR,we established the direct detection method from foods.
5.NaCl resistance : The relationship between the growth of L.monocutogenes and the NaCl concentration in broth was studied. At high concentration of NaCl, the growth was inhibited, but the virulence of L.monocytogenes for mouse mostly same. At the venetral injection, we observed the change of virulence.
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