贵州地方黄牛GHR基因遗传变异、功能预测及其与生长性状的关联性分析

doi:10.16431/j.cnki.1671-7236.2021.03.017

摘要/Abstract

摘要： 为探究黄牛生长激素受体（GHR）基因多态性，筛选出对贵州地方黄牛生长性状有显著影响的SNPs位点，本研究以150头贵州地方黄牛（关岭牛、思南牛、威宁牛各50头）为研究对象，以GHR为候选基因，根据GenBank收录的黄牛GHR基因外显子序列设计引物，提取贵州地方黄牛血液基因组DNA并构建混池；通过PCR扩增测序法验证GHR基因SNPs和分型，运用DNAStar、SPSS 19.0软件对GHR基因SNPs进行遗传学分析，并与贵州地方黄牛生长性状进行关联性分析，寻找各位点不同基因型间贵州地方黄牛生长性能差异，同时使用生物信息学分析GHR突变前后mRNA二级结构的变化，预测分析贵州地方黄牛GHR蛋白的结构与功能。结果显示，贵州地方黄牛GHR基因共检测出8个SNPs，分别为Exon9-C162T、Exon9-C201T、Exon9-G243C、Exon9-G383A、Exon9-A495T、Exon9-C622T、Exon9-C642T和Exon9-A650C，其中思南牛无Exon9-G243C。遗传学分析表明，除Exon9-G243C位点在威宁牛中偏离Hardy-Weinberg平衡外，其余SNPs均未偏离Hardy-Weinberg平衡。相关性分析发现，GHR基因突变位点在关岭牛和思南牛中均未检测到显著性差异，但在威宁牛中，Exon9-G243C基因型GC个体胸围显著低于GG和CC基因型（P<0.05），Exon9-A650C基因型AA个体的体斜长、坐骨端宽均显著高于CC基因型（P<0.05），而胸深显著低于AC和CC基因型（P<0.05），其余6个SNPs差异均不显著（P>0.05）。生物信息学分析发现突变前后，除Exon9-G383A mRNA二级结构未发生改变外，其余SNPs mRNA二级结构均发生了改变，Exon9-G243C、Exon9-A650C会影响蛋白质二级结构的变化，但突变不影响蛋白质三级结构的变化。此外，贵州地方黄牛GHR蛋白是一种分泌蛋白，具有信号肽剪切位点且具有6个N-糖基化位点，证实该基因变异程度高。本研究提示GHR基因Exon9-G243C、Exon9-A650C这2个SNPs对贵州地方黄牛生长性状具有显著影响，可作为贵州地方黄牛生长发育的候选分子标记。

关键词: 贵州地方黄牛; GHR基因; SNPs; 生长性状

Abstract: In order to analyze the polymorphism of GHR gene in cattle,SNPs with significant effects on the growth traits of native cattle in Guizhou were selected.This study used 150 Guizhou local Yellow cattle (Guanling cattle,Sinan cattle,Weining cattle each 50) as the research object,and GHR as the candidate gene,design primers based on the exon sequence of the Yellow cattle GHR gene included in GenBank.The blood of Guizhou local Yellow cattle was collected,blood genomic DNA was extracted and a mixed pool was contructed.The SNPs and typing of GHR gene was verified by PCR amplification sequencing method.Using DNAStar and SPSS 19.0 software to conduct genetic analysis on the SNPs of GHR gene and the correlation analysis with the growth traits in Guizhou local Yellow cattle,looking for the difference in the growth performance of Guizhou local Yellow cattle with different genotypes.At the same time,bioinformatics was used to analyze the changes of mRNA secondary structure before and after GHR mutation,predictive analysis of the structure and function of GHR protein in Guizhou local Yellow cattle.The results showed that a total of 8 SNPs were detected of GHR gene in Guizhou local Yellow cattle,namely Exon9-C162T,Exon9-C201T,Exon9-G243C,Exon9-G383A,Exon9-A495T,Exon9-C622T,Exon9-C642T and Exon9-A650C.Among them,Sinan cattle had no Exon9-G243C.Analysis of genetic characteristics showed that except for Exon9-G243C locus deviated from Hardy-Weinberg equilibrium in Weining cattle,the remaining SNPs did not deviate from Hardy-Weinberg equilibrium.Correlation analysis showed that GHR gene mutation sites were not detected in Guanling cattle and Sinan cattle.However,in Weining cattle,the chest circumference of individuals with Exon9-G243C GC genotype was significantly lower than that of GG and CC genotypes (P<0.05),the obliquee body length and ischial width of individuals with Exon9-A650C AA genotype were significantly higher than those with CC genotype (P<0.05),while the chest depth was significantly lower than that of AC and CC genotypes (P<0.05).The remaining 6 SNPs were not significant (P>0.05).Bioinformatics analysis showed that before and after the mutation,except for Exon9-G383A mRNA secondary structure did not change,the other SNPs mRNA secondary structure changed,Exon9-G243C and Exon9-A650C would affect the changes of protein secondary structure,but the mutation did not affect the change of the tertiary structure of the protein.In addition,Guizhou local Yellow cattle GHR protein was a secreted protein with signal peptide cleavage sites and 6 N-glycosylation sites.It showed that GHR gene had a high degree of variation.The results showed that the two SNPs of GHR gene Exon9-G243C and Exon9-A650C had significant effect on the growth traits of Guizhou local Yellow cattle,and might be used as candidate molecular markers for the growth and development of Guizhou local Yellow cattle.

Key words: Guizhou local Yellow cattle; GHR gene; SNPs; growth traits

中图分类号:

S827

周志楠, 吴雨, 陈祥, 宋汝谋, 陈伟. 贵州地方黄牛GHR基因遗传变异、功能预测及其与生长性状的关联性分析[J]. 中国畜牧兽医, 2021, 48(3): 932-945.

ZHOU Zhinan, WU Yu, CHEN Xiang, SONG Rumou, CHEN Wei. Genetic Variation and Function Prediction of GHR Gene and Correlation Analysis with Growth Traits in Guizhou Local Yellow Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 932-945.

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