基础医学与临床 ›› 2009, Vol. 29 ›› Issue (9): 924-928.

• 研究论文 • 上一篇    下一篇

近交系小鼠体细胞核移植囊胚的微卫星DNA鉴定

覃敏 何敏 宣强 莫林键 吴华 莫曾南   

  1. 广西医科大学第一附属医院泌尿外科 广西医科大学泌尿科学研究所 广西医科大学第一附属医院泌尿外科 广西医科大学泌尿科学研究所
  • 收稿日期:2008-08-06 修回日期:2008-10-30 出版日期:2009-09-20 发布日期:2009-09-20
  • 通讯作者: 莫曾南

Microsatellite DNA analysis of inbred mouse somatic cell nuclear transfer blastocysts

Min QIN, Min HE, Qiang XUAN, Lin-jian MO, Hua WU, Zeng-nan MO   

  1. Department of Urology, Institute of Urology and Nephrology , The First Affiliated Hospital,Guang Xi Medical University Department of Urology, Institute of Urology and Nephrology , The First Affiliated Hospital,Guang Xi Medical University
  • Received:2008-08-06 Revised:2008-10-30 Online:2009-09-20 Published:2009-09-20
  • Contact: Zeng-nan MO

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。

关键词: 细胞核/移植, 疾病模型, 微卫星DNA, 鉴定

Abstract: Objective To identify the source of the somatic cell nuclear transfer blastocysts , microsatellite DNA assay was developed. Methods DNA isolation from SCNT blastocyst,donor BALB/c mouse, recipient C57BL/6 mouse and KM mouse are 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 genetically related to the respective recipient mouse. Furthermore sequences of SCNT blastocysts are diferent from the control KM mouse. Conclusion Our results prove that the nucleus of somatic cell nuclear transfer blastocysts come from somatic nucleus of donor BALB/c mouse.

Key words: cell nuclear/transfer, disease model, microsatellite DNA, Identification