Surveys for different gene expression in male and female early embryos

这项研究的目的是分离男性和女性植入前胚胎，这在过去是不可能的。该策略是利用在性染色体中整合绿色荧光基因的转基因动物，通过这一研究项目，证明通过发射绿色荧光可以非常准确地预测植入前胚胎的性别。在这种无创性分离的基础上，我们制备了1500枚植入前胚胎，提取它们的mRNA，构建了cDNA文库。针对雄性和雌性小鼠胚胎特异性消减文库，将纯化的二次PCR产物克隆到pUC碱基载体中。90%的白色菌落含有带有插入物的质粒。建立了4个96孔板，其中包含来自每个文库的单个克隆。虚拟Northern杂交分析证实了三个驾驶员(雌性)特异克隆的差异表达。对于测试者(雄性)特异克隆，三个克隆中有五个确认为阳性。这些基因的序列被弄清楚了，现在准备进行进一步的研究。

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.