| Project/Area Number |
25660007
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Science in genetics and breeding
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| Research Institution | Hiroshima University |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 四型分泌系 / ジーンターゲティング / 遺伝子水平伝達 / 四型分泌系（T4SS） / プラスミド / ジーンターゲティング技術の開発 / 酵母への簡便迅速遺伝子導入法 / 4型分泌系 / VirB/D4-T4SS / RP4-T4SS / pBBR plasmid / ジーンターゲティング法 / 生物界間接合 / 高等植物のジーンターゲティング / RK2-T4SS / oriT |
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| Outline of Final Research Achievements |
RP4 plasmid transfers itself into eukaryotic cells by a Type4 Secretion System (RP4-T4SS), which is encoded in the plasmid. We found that the transferred liner ssDNA re-circularized exactly in yeast, and based on this finding, we intended to establish a gene-targeting method in plants.
We could not improve its transfer efficiency to the level, which is sufficient for applying to gene-targeting. Instead, we found a plasmid, which transfers and re-circularized in plant cells by Ti plasmid encoded Type 4 Secretion System (VirB/D4-T4SS). By using the plasmid, we constructed targeting vectors and helper plasmids, which possessed genes for enhancing homologous recombination. In spite of combining these vectors and helper plasmids, we have not detected the gene-targeting phenomenon in plant cells.
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