运用PCR对小鼠植入前胚胎进行性别诊断

根据C57BL6小鼠Y染色体重复序列145C5的碱基顺序，设计并合成一对引物，运用PCR扩增昆明白小鼠入前胚胎卵裂球DNA，以确定其性别，共对108枚活检胚胎的相应卵裂球进行了性别诊断，获雄性胚46枚，雌性胚62枚，移植后分别获雄性仔鼠4只，准确率100％（4／4），雌性仔鼠9只，准确率70％（9／13），本研究结果表明小鼠Y染色体重复序列145C5的碱基顺序在C57BL6小鼠和昆明白小鼠中基本一致，为农牧业动物进行性别选择和运用PCR进行单基因病植入前遗传学诊断提供了方法学基础。

Sexing of Mouse Preimplantation Embryos Using Polymerase Chain Reaction

In order to determine the sex of mouse embryo, 1 or 2 blastomeres were biopsied from Kun ming white mouse preimplantation embryo at 4 8 cells stage. The gDNA of the single blastomere was abstracted. According to the base sequence of 145C5, a repititive sequence of Y chromosome of mouse C57BL6, a pair of primer were asigned and synthesized. The gDNA was amplified using these primers. 108 mouse preimplantation embryos were sexed via this technique. 46 male embryos and 62 female embryos were transfered into five pseudopreganant mothers respectively. 4 male litters and 9 female litters were obtained. The diagnosis positive rate was 100%(4/4) and 70% (9/13)respectively. The result of PCR indecated that there was no difference between the repititive sequence of Y chromosome in mouse C57BL6 and Kun ming white mouse. The technique developed in this study might be further used for preimplantation genetic diagnosis of single gene defects.