摩拉水牛和尼里-拉菲水牛PRKAA2基因SNPs检测及遗传多样性分析

摘要

In order to study the polymorphism of protein kinase AM P-activated catalytic subunit alpha 2(PRKAA2),the genome DNA of Murrah and Nili-Ravi buffaloes were used as templates to amplify exon 4 and intron 3 partial sequences of PRK A A2 gene,the SNPs were detected by routine sequencing,and the genetic diversity was analyzed.The results showed that there was 1 SNP(c.462 G>A)in exon 4 and 3 SNPs(IVS3.557 T>C,IVS3.560 C> T and IVS3.565 G>A)in the intron 3.The genetic diversity results showed that wild homozygote and heterozygote were more advantages than mutant homozygote at c.462 G> A,the mutation homozygous geno-types were non-dominant genotypes at IVS3.557 T>C and IVS3.560 C> T,and the heterozygous genotypes were dominant genotypes at IVS3.565 G>A.IVS3.565 G>A in Murrah buffaloes and c.462 G> A in Nili-Ravi buffaloes were in non-equilibrium.All of 4 SNPs in Murrah buffaloes were at moderate polymorphism,c.462 G>A and IVS3.557 T>C in Nili-Ravi buffaloes were at low polymorphism,and IVS3.560 C> T and IVS3.565 G> A were at moderate polymorphism. The heterozygosity of IVS3.557 T >C was lower in two buffalo populations.The genotype fre-quency and gene frequency of IVS3.565 G> A in Murrah buffalo and c.462 G> A in Nile-Rafi buffalo were imbalanced,the genetic variation of IVS3.557 T > C in Nile-Rafi buffalo was low, without a high selection potential.4 SNPs would be combined into 5 haplotypes,T-C-G-G type was the dominant haplotype in two buffalo populations.In conclusion,4 SNPs of PRKAA2 gene in Mur-rah and Nili-Rafi buffaloes could provide a reference for marker-assisted selection breeding.%为研究水牛蛋白激酶AMP活化的催化亚基α2(protein kinase AMP-activated catalytic subunit alpha 2, PRKAA2)基因多态性,本试验以摩拉水牛和尼里-拉菲水牛基因组DNA为模板,扩增 PRKAA2基因外显子4及内含子3部分序列,通过常规测序法检测其SNP并进行遗传多样性分析.结果发现,PRKAA2基因外显子4内存在1个SNP位点(c.462 G>A),PRKAA2基因内含子3部分序列存在3个SNPs位点(IVS3.557 T>C、IVS3.560 C> T和IVS3.565 G>A).经遗传多样性分析表明,在c.462 G>A位点的野生纯合型和杂合型比突变纯合型更有优势,IVS3.557 T>C和IVS3.560 C> T位点的突变纯合型为非优势基因型,IVS3.565 G> A位点杂合型为优势基因型.IVS3.565 G>A位点在摩拉水牛群体中处于 Hardy-Weinberg非平衡状态;c.462 G> A位点在尼里 -拉菲水牛群体中处于 Hardy-Weinberg非平衡状态.4个SNPs位点在摩拉水牛群体中均为中度多态;c.462 G>A、IVS3.557 T>C位点在尼里-拉菲水牛群体中为低度多态,IVS3.560 C> T、IVS3.565 G>A位点为中度多态.IVS3.557 T>C位点在两个水牛群体中杂合度较低.说明摩拉水牛IVS3.565 G> A位点和尼里-拉菲水牛c.462 G>A位点的基因型频率和基因频率遗传状态不平衡,尼里 -拉菲水牛群体中IVS3.557 T>C位点遗传变异小,选择潜力不高.4个多态位点可以构建5种单倍型,其中T-C-G-G是摩拉水牛群体和尼里 -拉菲水牛群体的优势单倍型.综上,本研究检测的摩拉水牛和尼里-拉菲水牛 PRKAA2基因上4个SNPs位点可为水牛标记辅助选择育种提供参考.