乌珠穆沁绵羊RIPK2基因多态性与生长性状的关联

摘要

Objective]Based on the results of the previous genome-wide association on the sheep meat traits, we studied the possibility of the RIPK2 gene to be a novel candidate gene for the sheep growth traits, in order to confirm the GWAS results, as well as to find new molecular markers related to the sheep growth traits.[Method]In the 343 Ujumqin sheep experiment group, 30 DNA samples with a fixed concentration were selected to construct the DNA pool, primers were designed to extend exons and 1 000 bp of upstream and downstream regulate regions. PCR products were tested and the objective strap were sequenced. The sequence peak maps were analyzed by DNAMAN and Chromas2 softwares, Sequenom Mass-array technology was used to genotype the detected SNP including the SNP obtained by previous GWAS. Haploview was used to construct haplotypes and analyze linkage disequilibrium of the polymorphic loci. Association analysis between the SNPs in RIPK2 gene and the growth traits were performed using SPSS 22.0 software.[Result] Four SNPs were detected. In exon 4 was a A5536G synonymous mutation rs01, there were three genotypes on this site and the frequencies of AA, AG and GG were 0.42,0.45 and 0.13 individually, the dominant allele was A with a frequency of 0.65. In exon 7 was a T8952C missen variation rs02 with three genotypes. The frequencies of TT, TC and CC genotypes were 0.82,0.16 and 0.02. T was the dominant allele and its frequency was 0.9. Rs05 was a T2836C mutation in exon 10 with only two genotypes TT and CC. The frequency of genotype TT was 0.25 as well as CC was 0.75. In the upstream regions a G5181C variation rs30 was detected with three genotypes GG, GC and CC. The frequency of heterozygous genotype GC was 0.50 and the other two genotypes CC, GG were 0.18 and 0.32 respectively. G was the dominant allele with its frequency was 0.58.The variation T4456C detected by GWAS was rs34 in this study and there were three genotypes. The frequency of TT genotype was 0.49, the frequency of TC was 0.37 and the frequency of CC was 0.14.T was the dominant allele with a frequency of 0.68. Theχ2 test indicated all the SNPs fit with Hardy-Weinberg equilibrium except rs34. Rs02 was in low polymorphic status (PIC<0.25) and the other three SNPs were in moderate polymorphic status (0.25