| Project/Area Number |
04454068
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用生物化学・栄養化学
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| Research Institution | Hokkaido University |
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Principal Investigator |
KAWAMOTO Shinichi Fac.of Agriculture, Hokkaido Univ.Instructor, 農学部, 助手 (20169775)
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| Co-Investigator(Kenkyū-buntansha) |
NAITO Satoshi Fac.of Agriculture, Hokkaido Univ.Associate Professor, 農学部, 助教授 (20164105)
石川 雅之 北海道大学, 農学部, 助手 (70192482)
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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 |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1992: ¥5,100,000 (Direct Cost: ¥5,100,000)
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| Keywords | Arabidopsis thaliana / Reporter gene / Cell fusion / DNA transfer / Saccharomyces cerevisiae / 酵母 / YACライブラリー / 染色体機能部位 |
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| Research Abstract |
We developed a polyethylene glycol (PEG)-mediated direct DNA transfer method from intact Saccharomyces cerevisiae spheroplasts into Arabidopsis thaliana protoplasts. To monitor the DNA transfer from yeast to plant cells, b-glucuronidase (GUS) reporter gene in which a plant intron was inserted was used as a reporter. This intron-GUS reporter gene on a 2 mm-based plasmid vector was not expressed in yeast transformants, while it expressed GUS activity when the plasmid DNA was introduced into plant cells. When a mixture of S.cerevisiae spheroplasts harboring the plasmid and A.thaliana protoplasts was treated with PEG and high pH-high Ca^<2+> solution, GUS activity was detected in the extract of the plant cells after a three-day culture. Moreover, the GUS gene expression was resistant to micrococal nuclease treatment before and during PEG treatment. From these results, we concluded that plasmid DNA can be directly transfered from intact yeast spheroplasts to plant protoplasts by a nuclease-resistant process, possibly by the cell fusion.
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