广西巴马小型猪presenilin-1基因克隆及多态性分析

Cloning and Polymorphism Analysis of presenilin-1 Gene from Guangxi Bama Mini-pig

本研究的目的是寻找广西巴马小型猪presenilin-1基因SNP位点,并确定各基因型频率。利用RT-PCR的方法扩增并克隆presenilin-1基因,通过测序和对比确定SNP位点,RFLP方法分析100头近交系和封闭群的广西巴马小型猪的基因组DNA,确定该突变位点的分布情况。结果显示,成功克隆广西巴马小型猪presenilin-1基因CDS区全长并确定T188C错义突变位点,SNP检测确定,在33头近交系中TT型占93.9%,TC型占6.1%;在67头封闭群中TT型占64.2%,TC型占35.8%,在封闭群和近交系中均未发现CC基因型。本研究成功验证了presenilin-1基因的SNP位点,为下一步研究presenilin-1基因T188C突变位点的功能奠定基础。

The aim of this study is to look for Bama mini porcine presenilin-1 gene SNP loci and to determine the genotype frequencies. Using RT-PCR amplification and cloning ofpresenilin-I gene, SNP loci were determined by sequencing and comparing. RFLP analysis on genomic DNA of inbred and close groups of 100 Bama mini-pig was conducted to determine the distribution of the mutation. Results showed that full length CDS region of Guangxi Bama mini porcine presenilin-1 gene was cloned and T188C missense mutation was determined. SNP testing determined TT-93.9% and TC-6.1% in 33 inbred, whereas TT-64.2% and TC-35.8% were determined in 67 closed group, both of the two groups did not find CC genotype. This study did successfully validate presenilin-1 gene SNP loci. This research might provide an important supporting for further analyzing the function ofpresenilin-1 gene with T188C mutation.