| Budget Amount *help |
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
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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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