利用DHPLC研究我国几个地方绵羊品种黑素细胞刺激素受体基因单核苷酸多态性

利用测序及变性高效液相色谱(DHPLC)研究了蒙古羊、哈萨克羊、滩羊和藏绵羊黑素细胞刺激素受体（MSHR）基因单核苷酸多态性（SNP）。结果表明，在扩增片断长度为415bp范围内存在一个T317C突变，DHPLC可检测到该突变并被证明是一种高通量且简便的筛选方法。通过两次DHPLC可确定两个杂合子和一个纯合子，第一次DHPLC可迅速检测出由于形成异源双链而呈肩峰的杂合子（TC），但不能区分两个均呈单峰的纯合子（TT或CC）。第二次DHPLC将未知纯合子与已知序列的纯合子混合后进行，通过判定单峰或肩峰即可推断未知纯合子的基因型。所研究的4个绵羊群体在该突变位点均处于Hardy-Weinberg平衡状态。蒙古羊与哈萨克羊较近的遗传亲缘关系以及滩羊与藏绵羊较近的遗传亲缘关系与线粒体DNA的研究结果一致。

Study on SNP of Melanocyte Stimulating Hormone Receptor Gene in Several Chinese Indigenous Sheep Breeds Using DHPLC

Single nucleotide polymorphism (SNP) of melanocyte stimulating hormone receptor (MSHR) gene was studied by sequencing and denaturing high performance liquid chromatography (DHPLC) in Mongolian sheep, Kazakstan sheep, Tan sheep and Tibetan sheep. The results showed that there is a mutation at position 317 (T317C) within the length of 415bp and the DHPLC is a high-throughput and simple method for screening this mutation. The heterozygote (TC) with shoulder peak could be detected quickly at the first time of DHPLC, and two homozygotes (TT or CC) could be discriminated easily through two times of DHPLC when each homozygous DNA was mixed with a known homozygous reference sample at the second time of DHPLC. All of the populations for this site are in Hardy-Weinberg equilibrium. The results also indicated that Mongolian sheep and Kazakstan sheep had close relationship, Tibetan sheep and Tan sheep had close relationship. The relationship among breeds was consistent with that of microsatellite DNA.