| Project/Area Number |
17590392
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
KOMATSUZAWA Hitoshi Hiroshima University, Graduate School of Biomedical Science, Associate Professor, 大学院医歯薬学総合研究科, 助教授 (90253088)
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| Co-Investigator(Kenkyū-buntansha) |
SUGAI Motoyuki Hiroshima University, Graduate School of Biomedical Science, Professor, 大学院医歯薬学総合研究科, 教授 (10201568)
FUJIWARA Tamaki Hiroshima University, Graduate School of Biomedical Science, Research Associate, 大学院医歯薬学総合研究科, 助手 (90274092)
OHARA Masaru Hiroshima University, Graduate School of Biomedical Science, Research Associate, 大学院医歯薬学総合研究科, 助手 (80253095)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Staphylococcus aureus / antibiotic resistance / sugar metabolism / Microarray / ribozyme |
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| Research Abstract |
We investigated the interaction of sugar metabolism with the susceptibility to vancomycin and methicillin in Staphylococcus aureus, and had several findings described below.
1) Construction of PTS mutants and characterization of these mutants : We found possible 17 PTS systems such as specific PTS for glucose and for fructose, so we constructed 17 PTS mutants. We measured the susceptibility to antibiotis, and found no significant difference between the mutant and the parent.
2) Construction of ccpA-mutants and its characterization: We constructed the mutant of ccpA, which is involved in global regulation of glucose metabolism. CcpA-mutant reduced the resistance level to methicillin. From the microarray analysis, we found the genes regulated by CcpA.
3) Analysis of GlmS-ribozyme: To know the critical regions for ribozyme activity in glmS-mRNA, we constructed the various length of DNA fragment containing promoter region, and cloned these fragments in one vector for XylE-reporter analysis. As a result, the region responsible for ribozyme activity is about 300 by region upstream of glmS-orf. To evaluate the ribozyme activity, we set up the method using real-Time PCR, and found that glmS-mRNA was cleaved in specific sites by additon of N-acetylglucosamine, and also digested glmS-mRNA itself.
4) Microarray analysis of the expression profile by addition of various kinds of sugars: We analyzed the expression of all genes in S. aureus by addition of various kinds of sugars, and found specific PTS for each sugar. Also, we evaluated the specific genes affected by each sugar.
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