牦牛FSHR基因5′端部分序列克隆及生物信息学分析

Cloning and Sequence Analysis of 5' Flank Region of FSHR Gene in Yak

利用PCR和T-A克隆技术,测定了麦洼牦牛和杂种犏牛的FSHR基因5′端侧翼区和第1外显子的核苷酸序列,并运用BLAST、DNAMAN、Clustal等生物信息学软件与其他物种的相应序列进行了同源性比对分析,为牦牛FSHR基因结构、犏牛雄性不育、牦牛遗传多样性,以及FSHR基因与牦牛的繁殖、产肉、产奶性状相关分析提供了理论基础。结果表明:牦牛和犏牛FSHR基因5′端侧翼区长度为970bp,普通牛、羊和猪的该序列长度分别为970、973和981bp,序列长度差异较小;序列突变以碱基替换为主,牦牛与犏牛、普通牛、羊及猪的核苷酸序列一致性分别为99.4%、99.3%、98.2%、96.9%,一致性较高,为同源基因。聚类分析中牦牛与犏牛首先相聚,再依次与普通牛、羊、猪聚在一起。根据核苷酸序列推测,氨基酸组成分析显示,牦牛和犏牛的该蛋白疏水性,蛋白的亲水性和稳定性较差,为不稳定的脂溶性蛋白;二级结构主要以α螺旋和随机卷曲为主。

The complete sequences of FSHR 5＇ flank region （5＇ flank region and exon I ） of Maiwa yaks and Pianniu were determined by PCR （polymerase chain reaction） method and the T-A clone method, then the homologous analysis of the complete sequences of the FSHR 5＇ flank region between Maiwa yaks and Pianniu and other species was conducted through bioinformatics softwares such as BLAST, DNAMAN, Clustal, etc. , hoping to provide theoretical support for the future researches and studies on the gene structure of FSHR, Dzo male sterility, Yak genetic diversity, and correlation analysis on FSHR Gene and the breeding, meat production, and milk production traits of yaks. The research results showed that FSHR 5＇ flank region sequence length is 970 bp , and those of cattle, sheep and pigs are 970, 973 and 981 bp , only with slight difference in between. Sequence mutation was mainly in the form of base substitution; the nucleoside sequences homology of yak, pianniu , cattle, sheep and pig are 99. 4 %, 99. 3 %, 98. 2 %, 96. 9 % respectively. Yak has the closest relationship with pianniu followed by cattle, sheep, and then pig. According to the analysis of amino acid components predicted by the nucleoside sequence, FSHR protein in Yak and Pianniu is unstable, having poor hydrophobicity, hydrophilicity , and stability, and its secondary structure is mainly helix and randomly coiled.