Surveys for different gene expression in male and female early embryos

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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.