| Project/Area Number |
20580135
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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 |
Food science
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| Research Institution | Kyushu University |
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Principal Investigator |
HONJOH Ken-ichi Kyushu University, 大学院・農学研究院, 准教授 (00264101)
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| Co-Investigator(Kenkyū-buntansha) |
MIYAMOTO Takahisa 九州大学, 大学院・農学研究院, 教授 (70190816)
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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: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2009: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2008: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
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| Keywords | 食品保蔵学 / 耐凍性 / 植物 / 酸化ストレス |
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| Research Abstract |
NADPH-dependent thioredoxin reductase, which is localized in chloroplast, from Chlorella (CvNtrC) was confirmed to be low-temperature-inducible at transcriptional and translational levels. Peroxiredoxin (CvPrx), which works with CvNtrC in Chlorella, was isolated by in vitro pull-down assay and identified by N-terminal amino acid sequencing. The gene encoding CvPrx was cloned and its recombinant mature protein (mCvPrx) was produced in Escherichia coli by other researchers. We obtained the CvPRX gene and mCvPrx protein from the researchers. We also produced recombinant mature CvNtrC (mCvNtrC) protein in E. coli. The two proteins coordinately showed peroxidase activities against H_2O_2 and peroxides. Furtheremore, we transformed Saccharomyces cerevisiae with mCvNTRC and/or mCvPRX genes. Expression of mCvNTRC or mCvNTRC/mCvPRX led to enhancement of S.cerevisiae against freezing-, oxidative-, and heat-stress tolerances, suggesting that these two genes play important roles in stress tolerance. To make a knock-out strain of Chlorella for NTRC gene, we developed the transformation method for C.vulgaris C-27. Furthermore, we introduced the genes into Arabidopsis thaliana with the floral dip method.
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