| Project/Area Number |
08457600
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KAMO Naoki Hokkaido Univ.Fac.of Pharm.Sci.Prof., 薬学部, 教授 (10001976)
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| Co-Investigator(Kenkyū-buntansha) |
SUMI Masato Hokkaido Univ.Fac.of Pharm.Sci.Inst., 薬学部, 助手 (30281819)
NARA Toshifumi Hokkaido Univ.Fac.of Pharm.Sci.Inst., 薬学部, 助手 (30241350)
MIYAUCHI Seiji Hokkaido Univ.Fac.of Pharm.Sci.Lec., 薬学部, 講師 (30202352)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1997: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1996: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Keywords | Haloferax volcanii / ABC super family / Enteroccus hirae / Bacillus subtilis / doxorubicin / ethidium / glucose / hexokinase / Thermus thermophilus / MDR(multi-drug resistance) / ABC transporter / ATP binding casette(ABC) / PCR / Halobacterium volcaii / 多剤排出輸送体 / EmrE / Haloferax volcanii / 大腸菌 / metabolic stress / Enterococcus hirae / ロ-ダミン123 |
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| Research Abstract |
(1) A mutant of Haloferax volcanii resistant for doxorubicin was isolated and its resistance was proved to be caused by an efflux pump driven by ATP-hydrolysis.
(2) Wild-type cells of H.volcanii expressed this efflux pump, and its activity increased when cells were cultured in the presence of excess qlucose and amino acids.
(3) Partial DNA sequence was determined which may code this efflux pump, and had high similarity with that of ABC transporters.
(4) A mutant of Enoterococcus hirae was isolated which showed high resistance against ethidium bromide. By shot-gun cloning using E.coli as a hast cell, a factor conferring the resistance was 165 rRNA.The fact that 165 rRNA is related to the resistance is a new finding.
(5) An efflux activity of Bacillus subtilis ISW1214 was decreased when cultured in the presence of glucose.When non-metacolic sugars were used instead of glucose, this phenomena were not found. Detailed analysis showed a good negative correlation between intracellular concentration of fructose 6-phosphate and the efflux activity.
(6) We cloned a gene coding a requlator protein that controls an expression of the efflux transporter in Bacillus subtilis ISW1214. It is quite interesting that a part of this gene has high similarity with a part of hexokinase, and enzyme of qlucose metabolism.
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