Bent promoter of epsilon-toxin gene from Clostridium perfringens : analysis and application to protein expressions.
| Project/Area Number |
20590448
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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 | Kagawa University |
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Principal Investigator |
MIYATA Shigeru Kagawa University, 医学部, 講師 (90314913)
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| Co-Investigator(Kenkyū-buntansha) |
NARIYA Hirofumi 香川大学, 医学部, 助教 (30452668)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2010: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2009: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2008: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
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| Keywords | ウェルシュ菌 / イプシロン毒素 / bent DNA / プロモーター / フェレドキシン / 糖鎖分解酵素 / クロストリパイン / AT-rich遺伝子 / コラゲナーゼ / NagH |
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| Research Abstract |
Phased A-tracts form a bent structure of DNA. We found a novel type of bent DNA structure, which stimulates the gene transcription, downstream of a Clostridium perfringens epsilon-toxin promoter. When we compared the temperature dependency of gene transcription from epsilon-toxin promoter to those from other bending promoters, alpha-toxin and ferredoxin promoters ; epsilon-toxin, alpha-toxin and ferredoxin promoters exhibit high-level transcription at 37℃, 25℃ and over a wide rangeof temperature, respectively. Thus, we constructed a novel expression vector using ferredoxin promoter to express efficiently large clostridial proteins in C.perfringens. Our expression system was successfully applied to produce large amounts of ColH, NagH, NagJ, NanJ, and BgaA, which are poorly expressed in Escherichia coli expression system. These proteins were purified in sufficient amounts from the supernatants of the recombinant C.perfringens cultures.
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Report
(4 results)
Research Products
(12 results)
