牛SLC27A1基因SNPs筛选及其与中国荷斯坦奶牛产奶性状的关联分析

目的]对牛SLC27A1基因进行SNPs筛选并与中国荷斯坦奶牛的产奶性状进行关联分析,以期发现对中国荷斯坦奶牛产奶性状有显著影响的SNP位点。方法]根据性能测定记录选取48头中国荷斯坦奶牛,提取血样DNA,组成2个DNA池,用于SNPs筛选,采用PCR-SSCP和克隆测序方法对DNA池进行SLC27A1基因SNPs筛选。针对发现的SNP位点,采用PCR-RFLP方法对另外231头中国荷斯坦奶牛进行群体基因型检测,采用SAS（8.02）GLM过程对各基因型与产奶性状进行关联分析。结果]在exon3发现T112C位点,在3‘UTR发现G64A位点,T112C为同义突变;经PCR-RFLP检测,发现在T112C位点存在TT、TC、CC3种基因型,G64A位点存在GG、GA、AA3种基因型。2个位点均处于Hardy-Weinberg平衡状态。T112C位点的CC型个体的产奶量极显著高于TC型个体（P〈0.01）,3种基因型对乳蛋白率和乳脂率影响均不显著（P〉0.05）,乳蛋白率呈现CC〉TC〉TT的趋势,乳脂率呈现TT〉TC〉CC的趋势;G64A位点3种基因型对产奶量、乳蛋白率、乳脂率的影响均不显著（P〉0.05）,但产奶量呈现GA〉GG〉AA的趋势,乳蛋白率和乳脂率呈现AA〉GG〉GA的趋势。结论]该基因T112C位点与产奶量性状有一定的关联性,通过提高CC基因型频率有望提高中国荷斯坦奶牛的产奶量,SLC27A1基因可作为调控中国荷斯坦奶牛产奶量的候选基因,同时为该基因的标记辅助育种和进一步研究奠定了良好基础。

Identification of SNPs in Bovine SLC27A1 Gene and Its Effects on Milk Production Traits in Chinese Holstein Cattle

Objective] This study discussed the SNPs of SLC27A1 gene and its relationship with milk production traits in Chinese Holstein Cattle in order to find the SNP site which had significant effect on milk production traits in Chinese Holstein Cattle. Method] DNA was extracted from the bleed of 48 Chinese Holstein Cattle selected according to phenotypic character and mixed into DNA pool for SNPs detection by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and cloning sequencing. Then different genotypes were detected in other 231 Chinese Holstein Cattle by PCR-RFLP. The association between genotype and production traits was assessed by GLM procedure,SAS version 8.02. Result] There were T112C in exon3 and G64A loci in 3'UTR,among them the T112C in exon3 was synonymous mutation. There were 3 genotypes TT,TC and CC in T112C locus and 3 genotypes GG,GA and AA in G64A locus. The population was at Hardy-Weinberg equilibration. Cows with genotype CC had significantly highest milk yield than those with genotype TC (P< 0.01),and there were no significant differences among the 3 genotypes on milk protein percent and milk fat percent (P > 0.05),but the tendency of CC > TC > TT on milk protein percent and the tendency of TT > TC > CC on milk fat percent were showed. There was no significant difference among the 3 genotypes of G64A loci on milk yield,milk protein percent and milk fat percent (P > 0.05),but the tendency of GA > GG > AA on milk yield and the tendency of AA > GG > GA both on milk protein percent and milk fat percent were showed. Conclusion] There was certain relation between the T112C locus and milk yield traits; It may improve milk yield to raise the frequency of genotype CC; SLC27A1 gene could be a useful candidate gene in selection program on milk yield traits in Chinese Holstein Cattle,which provided a theoretical basis for the marker-assisted breeding and further study of SLC27A1 gene.