2011 Fiscal Year Final Research Report
Mutation burst by conflict of two mouse subspecies genomes
| Project/Area Number |
23657009
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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 |
Genetics/Genome dynamics
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| Research Institution | National Institute of Genetics |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
TAMURA Masaru 国立遺伝学研究所, 系統生物研究センター, 助教 (50370119)
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| Project Period (FY) |
2011
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| Keywords | ゲノム構築 / 再編 / 維持 |
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| Research Abstract |
We carried out two experiments to elucidate mechanism by which high-frequency spontaneous mutations(Recombination-induced-mutation : Rim) is induced in mice with recombinant MHC haplotypes between two mouse subspecies. First, we tried to detect genome alterations similar to Rim by PCR-based monitoring of Rim-type mutations using pooled sperm DNAs of intra-MHC recombinant mice. As a result, we could not observe expected PCR-products by this method. One possibility to explain this result is that there are different types of mechanism for induction of Rim. Alternatively, present intra-MHC recombinant mice have lost the activity to give rise to Rim. Second, we tried to develop an efficient assay system to detect DNA-double strand breaks and subsequent recombination for single male mouse. To do this, now we constructed the DNA construct that consists of recombination-target DNA sequence for Prdm9 protein, which is known to determine recombination site by introducing Histon3 chemical modification as hotspot mark, and of GFP reporter connected to the Prdm9 target sequence. Now we are introducing this construct into mouse genome by means of Tol2 transposon system.
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