【摘要】 目的利用微卫星DNA技术进行体细胞核移植囊胚的鉴定。方法根据Mouse Genome Database设计小鼠微卫星DNA多态聚合酶链反应引物,提取近交系小鼠体细胞核移植囊胚、供体BALB/c小鼠、受体C57BL/6小鼠及昆明小鼠的基因组DNA,使用巢式聚合酶链反应扩增4个微卫星位点DNA片段,即D3Mit28、D11Mit258、D12Mit136及D14Mit50。对反应产物进行琼脂糖凝胶电泳验证。结果巢式聚合酶链反应可扩增微量基因组DNA,通过微卫星DNA序列的扩增,证明核移植囊胚的微卫星DNA与供体细胞完全相同,而与受体细胞或者对照细胞无亲缘关系。结论体细胞核移植囊胚基因组来源于供体BALB/c小鼠。更多还原

【Abstract】 Objective To identify the source of the somatic cell nuclear transfer blastocysts with microsatellite DNA technology.Methods DNA was isolated from SCNT blastocyst,donor BALB/c mouse,recipient C57BL/6 mouse and KM mouse were tested by nested polymerase-chainreaction(PCR) analysis.Primers designed for four specific microsatellite locus(D3Mit28,D11Mit258,D12Mit136,D14Mit50)were employed.The amplified PCR products from the final reaction was analyzed by agarose gel electrophoresis and visualized with ethidium bromide staining.Results Amplification of genomic material by nest PCR represents the most sensitive method for the detection and might be detected successfully even though very low DNA copies.Microsatellite DNA analyses examining four loci confirm that all the SCNT blastocysts were genetically identical to the donor mouse.Additionally,the SCNT embryos are not related to the respective recipient mouse.Furthermore sequences of SCNT blastocysts are diferent from the control KM mouse.Conclusion The nuclei of somatic cell nuclear transfer blastocysts come from somatic nucleus of donor BALB/c mouse.更多还原