| Project/Area Number |
11480220
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | Osaka University |
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Principal Investigator |
OKABE Masaru Osaka University, Genome Information Research Center, Professor, 遺伝情報実験センター, 教授 (30089875)
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| Co-Investigator(Kenkyū-buntansha) |
伊川 正人 大阪大学, 遺伝情報実験施設, 助手 (20304066)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2001: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 2000: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1999: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | sex differentiation / sex chromosome / GFP / X chromosome / sexing of embryos / トランスジェニックマウス / 雌雄分別 / ディファレンシャルディスプレイ |
|---|
| Research Abstract |
The purpose of this study is to separate male and female preimplantation embryos, which was not possible in the past. The strategy was to employ the transgenic animals that integrated green fluorescent gene in their sex chromosomes.
Through this research project, it was proved that the sexes of preimplantation embryos are very accurately predicted by the emission of green fluorescent light. Based on this non-invasive separation, we have prepared 1500 each male and female preimplantation embryos and their mRNA was extracted to make cDNA library. For male and female mouse embryo-specific subtracted cDNA pools, purified PCR-amplified secondary PCR product was cloned into the PUC base vector. The 90% of white colonies contain plasmid with insert. Four 96-well plates containing individual clones from each library were established. Virtual northern blot analysis confirmed differential expression of three Driver (female)-specific clones.
For Tester (male)-specific clones, five out of three clones are confirmed positive. The sequences of these genes are clarified and now ready for the further investigations.
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