2014 Fiscal Year Final Research Report
Development of one-step gene disruption system of diploid shochu yeast strain using loss of heterozygosity
| Project/Area Number |
24580118
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Kagoshima University |
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Principal Investigator |
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Keywords | ヘテロ接合性消失 / 焼酎酵母 / ２倍体 / 遺伝子破壊 |
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| Outline of Final Research Achievements |
It has been thought to be difficult to obtain mutant strain, because shochu yeast is diploid. Thus mutation in one of the two allelic genes is hard to appear as a mutant phenotype. In this study, we constructed a gene disruption system of diploid shochu yeast by using LOH (Loss of Heterozygosity). We designed and constructed a gene disruption cassette by combination of LYS5 a gene involved in Lysine biosynthesis and URA3 a gene involved Uracil biosynthesis. By using this gene disruption cassette, we succeeded in single gene disruption of LEU2 a gene involved in Leucine biosynthesis, and thereafter obtained a double gene disruptant by inducing LOH. Further, we succeeded second round gene disruption after removing the gene disruption cassette from LEU2 double gene disruptant.
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| Free Research Field |
応用微生物学
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