甘南牦牛Lfcin基因克隆及序列分析

Clone and sequence analysis of Lfcin gene of the Gan Nan Yak's

利用PCR技术从甘南牦牛基因组DNA中扩增获得了含乳铁蛋白素(lactoferricin,Lfcin)基因的DNA序列,将该DNA序列连接于pGEM-Teasy载体并测序,将甘南牦牛含Lfcin基因的DNA序列与奶牛相应序列进行比对,同时对牦牛、人、小鼠等物种的Lfcin蛋白进行序列分析和进化树分析。结果:试验首次克隆获得了含甘南牦牛乳铁蛋白(lactoferrin,LF)第二外显子的DNA序列(在Gen-Bank中获得登陆号EU247256),共778 bp,其中LF基因第二外显子长165 bp,编码55个氨基酸,Lf-cin蛋白占25个氨基酸;序列分析结果显示,克隆获得的牦牛DNA序列与奶牛相应序列存在3个碱基的变异;牦牛和奶牛的Lfcin蛋白质序列完全相同;各物种Lfcin蛋白具有较高的同源性,Lfcin蛋白进化树符合物种进化规律。

The Lfcin（Lactoferricin） gene was amplifed by PCR. The gene was cloned to pGEM -T is vector, and the product was sequenced . Gan Nan yak and cow＇ s Lfcin sequences were similarity. Yak, human, mouse＇ s Lfcin protein sequences and the phylogenetic tree of Lfcin protein in different species were analysed. The DNA sequence that contains the Lfcin gene of Gan Nan yak was cloned, constituted by 778 bp, Lfcin coding region that code 25 amino acids being 75 bp length. According sequence analysis, there are 3 different bases between cow and yak＇ s sequence. Yak and cow＇ s Lfcin gene code same sequences and there were high homology in different species. The phylogenetic tree of different species Lfcin protein accords with the order of evolution.