贵州杂交黄牛MSTN基因cDNA克隆、生物信息学及多态性分析  

cDNA Cloning,Bioinformatics and Polymorphism Analysis of MSTN gene of Hybrid Guizhou Yellow Cattle

作　　者：王可人 游绍航 主性[1] 田兴贵[1] WANG Keren;YOU Shaohang;ZHU Xing;TIAN Xinggui(College of Animal Science,Guizhou University,Guiyang,Guizhou 550025,China;Tongren Wanshan Agriculture and Rural Affairs Bureau,Tongren,Guizhou 554200,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]铜仁市万山区农业农村局,贵州铜仁554200

出　　处：《家畜生态学报》2023年第4期13-21,共9页Journal of Domestic Animal Ecology

基　　金：贵州本地黄牛MSTN基因多态性与生长性能分析(贵大人基合字(2017)20号);贵州省科技创新人才团队项目(黔科合平台人才2019-5615)。

摘　　要：为比较贵州杂交黄牛肌肉生长抑制素(Myostatin,MSTN)性质、结构和多态性与本地黄牛的差别,该研究先通过RT-PCR克隆MSTN基因cDNA并进行生物信息学分析,再通过PCR-SSCP结合直接测序法筛查贵州本地黄牛和杂交黄牛MSTN基因外显子多态性。结果在贵州杂交黄牛MSTN基因CDS区序列发现4个突变位点,其中3个为错义突变:76 bp处G/A替换,引起天冬酰胺变为天冬氨酸;79 bp处G/A替换,引起丝氨酸变为甘氨酸;1066 bp处G/A替换,引起谷氨酸变为赖氨酸。267 bp处G/A替换与525 bp处A/G替换,为同义突变。MSTN蛋白基本理化性质和结构与普通牛基本一致,但有细微的差别;通过基因同源性和系统进化树,结合motif、蛋白保守域和结构域分析,可知MSTN基因在哺乳动物中高度保守,但也存在遗传多样性。贵州本地黄牛外显子发现3个SNP位点,分别为g.6279107 G>C、g.6281238 T>C和g.6283933 A>C,贵州杂交黄牛中未发现多态位点。单倍型分析共发现6种单倍型和14种双倍型,双倍型H3H5频率最高,且杂交牛的双倍型均为H3H5,之后可一步将此与生长性状进行关联分析研究。In order to compare the character,structure and polymorphism of myostatin(Growth differentiation factor,MSTN)between hybrid Guizhou yellow cattle and local cattle,the cDNA of MSTN gene of hybrid Guizhou yellow cattle was firstly cloned by RT-PCR and analyzed by bioinformatics.Then,the single nucleotide polymorphism site(SNP)of MSTN gene exon was screened by PCR-SSCP combined with direct sequencing method.The results showed that there were five mutation sites in the CDS region of MSTN gene between hybrid Guizhou yellow cattle and Bos taurus,the G/A substitution at 76 bp resulted in the change from asparagine to aspartic acid,the G/A substitution at 79 bp from serine to glycine,and 1066 bp G/A substitution from glutamic acid to lysine,but G/A substitution at 267 bp and A/G substitution at 525 bp were synonymous mutations.The basic physical and chemical properties and protein structure of MSTN gene are basically the same as those of Bos taurus,but there were subtle differences.The homology and phylogenetic tree analysis,combined with the motif,conserved domain and structural domain analysis,indicated that MSTN gene was highly conserved in mammals,but there was genetic diversity.Three SNPs were found in exons of Guizhou yellow cattle,which were g.6279107 G>C,g.6281238 T>C,and g.6283933A>C,but no polymorphic loci were found in hybrid Guizhou yellow cattle.In haplotype analysis,a total of six haplotypes and fourteen diplotypes were found.Diplotype H3H5 had the highest frequency,and all the diplotypes of hybrid Guizhou yellow cattle were H3H5.It is,therefore,necessary to analyze the association between diplotype and growth traits of yellow cattle to verify whether it is an important reason affecting the growth traits of yellow cattle.

关 键 词：贵州杂交黄牛 肌肉生长抑制素基因 基因克隆 生物信息学 多态位点 

分 类 号：S823.2[农业科学—畜牧学]