| Project/Area Number |
04806017
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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 |
応用微生物学・発酵学
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| Research Institution | University of Osaka Prefecfure |
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Principal Investigator |
SAKAI Takuo University of Osaka Prefacture College of Agriculture Professor, 農学部, 教授 (50081500)
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| Co-Investigator(Kenkyū-buntansha) |
KISHIDA Masao University of Osaka Prefacture College of Agriculture Assistant Professor., 農学部, 助手 (90211193)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | yeast / mitochondria / molecular breeding / apocytochrome b / ミトコンドリアDNA / 遺伝子 |
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| Research Abstract |
We attempted to develop new method to introduce heterogeneous gene to yeast cell. In the method, heterogeneous gene is introduced to mitochondria DNA (mtDNA) using integration vecter plasmid on which heerogeneous gene is conected with marker gene. As the marker, we selected apocytochrome b (Cob) , wich is an enzyme on respiration chain of the yeast, and Cob-deficient yeast lose its respiration ability. The enzyme has relatively low molecular weight and its gene seemed to be suitable for the gene marker. If the gne connected with Cob gene is introded on Cob deficient mtDNA,the respiration ability is restore so as to be found introduction of new gene.
We constructed the above gene introduction system, according the following steps
(1) Development of effective method to prepare mitochondria and its introduction to yeast cells.
(2) Development of effective method to prepare mtDNA.
(2) Cob gene cloning by PCR method.
In the next step, We intend to introduce Lac Z gene in Cob gene to induce incomplete Cob gene, then, introduce it on mtDNA resulting to introduce Cob deficent mutant.
Finally, construction of "Cob deficient cell-Cob harboring vecter system" is accomplished.
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