| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Research Abstract |
Arabidopsis tester lines possessing an I-SceI recognition site and an inducible I-SceI expression system were established for monitoring protein attachment at DNA double strand break (DSB) site and for monitoring gene targeting events. By using this experimental material, it was revealed that gene targeting efficiency was increased by a factor of 5-10 by DSB induction during transformation of gene targeting vector.
It is known that T-DNAs preferentially integrate at DNA DSB sites. Previous studies show that VirD2, which is one of the key Agrobacterium tumefaciens proteins involved in T-DNA processing and transfer is important for efficient T-DNA integration in to the plant genome. However there is no direct evidence to show the role of VirD2 in T-DNA integration step. For detecting in vivo VirD2 attachment at DSB sites, I established chromatin immunoprecipitation assay using protoplast of this Arabidopsis tester line with transfection of VirD2 expression cassette and DSB induction.
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