高山美利奴羊INHA基因多态性生物信息学分析及与产羔数关联分析

Polymorphism and Bioinformatics Analysis of INHA Gene and Its Association with Litter Size in Alpine Merino Sheep

本研究旨在研究高山美利奴羊INHA基因外显子多态性及其与产羔数的相关性。本实验通过比对高山美利奴羊全基因组测序结果中不同个体INHA外显子,分析高山美利奴羊INHA基因的单核苷酸多态性,应用生物信息学软件分析高山美利奴羊INHA基因突变前后不同等位基因的m RNA二级结构、蛋白质的二级结构及三级结构。通过上述分析,将引起高山美利奴羊INHA编码氨基酸变化的位点作为该基因的特异性候选位点,通过直接测序法分析高山美利奴羊特异性候选位点INHA基因多态性,并分析其与产羔数的相关性。结果发现,高山美利奴羊INHA基因外显子区域存在3个SNPs,分别为T206A (Met→Lys)、T387A(Thr→Thr)、G900A (Pro→Pro);INHA基因3个SNPS都改变了RNA的最小自由能以及二级结构,错义突变T206A (Met→Lys)引起蛋白质二级结构和三级结构的改变;高山美利奴羊INHA基因T206A突变表现出3种基因型分别命名为TT、TA、AA,基因型与产羔数关联分析发现TT、TA基因型个体的产羔数极显著高于TT基因型个体(p<0.01)。本研究初步表明INHA基因是影响高山美利奴羊产羔数的一个主效基因。

The aim of this study was to investigate the polymorphism of the INHA gene exon in Alpine Merino sheep and its correlation with litter size. In this experiment, we analyzed the single nucleotide polymorphism of the INHA gene of Alpine Merino sheep by comparing the INHA exons of different individuals in the whole genome sequence of Alpine Merino. The bioinformatics software was used to analyze the m RNA secondary structure,secondary structure and tertiary structure of different alleles of Alpine Merino sheep. Through the above analysis,the site that caused the amino acid change of the INHA gene of the Alpine Merino sheep was used as a specific candidate site for the gene, The gene polymorphism of INHA in the Alpine Merino sheep-specific candidate locus was analyzed by direct sequencing, and its correlation with litter size was analyzed. The results showed that there were three SNPs in the exon region of the INHA gene of Alpine Merino, which were T206 A(Met→Lys), T387 A(Thr→Thr) and G900 A(Pro→Pro). Further analysis revealed that each mutation site caused changes in RNA secondary structure and minimum free energy. The missense mutation site T206 A(Met→Lys) caused changes in protein secondary structure and tertiary structure. The T206 A mutation of INHA gene in Alpine Merino sheep showed three genotypes, named TT, TA and AA respectively. The correlation analysis between genotype and litter size showed that the number of litters in TT and TA genotypes was significantly higher than that of TT genotypes(p<0.01). This study initially showed that INHA was a major gene affecting the litter size of Alpine Merino sheep.