| Project/Area Number |
63560114
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
発酵・醸造
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| Research Institution | Osaka Prefecture University |
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Principal Investigator |
SAKAI Takuo University of Osaka Prefecture Department of Agricultural Chemistry Professor, 農学部, 教授 (50081500)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥100,000 (Direct Cost: ¥100,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Saccharomyces cerevisiae / oversecretion of enzyme / expression of heterologus gene / CDC mutant / Calcium ion / temperature sensitive mutant / 酵母の形質転換 / G2アレスト / 温度感受性の酵母 / カルシウムイオン / 酵母の遺伝子組換え / 異種遺伝子高発現酵母 / 遺伝子の発現とカルシウム / 遺伝子の組換え / プロトペクチン可溶化酵素 / 酵母のプロモーター |
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| Research Abstract |
The research on breeding of efficient strains to produce heterologous gene product have been done. A mutant responsible for an overproduction phenotype on heterologous gene product in S. cerevisiae has been isolated, using the gene coding for polygalacturonase (PG) from KZLwvem"s marxianus as a marker.
The results obtained are as follows :
(1) A mutant responsible for an oversecretion of PG was obtained. The mutant (SSM5-2) produced PG by 10- to 30-fold in parent strain.
(2) Contents of lipids in the cell membrane of the mutant was different from those of parent strain, and the conditions for preparing competent cell were also different from parent strain. A simple and efficient method for transformation with plasmids was established.
(3) SSM5-2 was temperature-sensitive lethal mutant and it growth was arrested with the production of PG, and it was confirmed to be G2 arrest. The G2 arrest was found to be recovered by Ca^<2+>.
(4) A gene which promote the PG production (PGPPF) was found on upstream region of PG gene of K. marxianus. The base sequence of PGPPF was determined.
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