山羊ZBP1基因多态性及与产羔性能的关联分析

Association Analysis Between Polymorphisms of ZBP1 Gene and Lambing Performance in Goats

【目的】试验旨在分析山羊Z-DNA结合蛋白(Z-DNA binding protein 1,ZBP1)基因3′-端非翻译区(3′-UTR)、内含子1和外显子7区域中单核苷酸多态性(single nucleotide polymorphism,SNP)位点与产羔性能之间的关联性,为高繁殖力山羊分子育种提供新的遗传标记。【方法】选取麻城黑山羊、黑头羊和波尔山羊作为研究样本,采集血样提取DNA,根据转录组数据所选SNP位点在山羊ZBP1基因序列中的位置信息设计引物,利用多重单碱基延伸SNP分型技术(SNaPshot)分析所选SNP位点的遗传多样性,采用一般线性模型(GLM)对ZBP1基因SNP位点与3个品种山羊的产羔性能进行关联分析。【结果】山羊ZBP1基因中存在12个潜在的SNPs位点,其中7个SNPs位点(g.9734 A>G、g.9772 T>C、g.352 C>T、g.955 C>T、g.1880 G>A、g.2566 T>C和g.7919 G>A)具有多态性,除g.9734 A>G在麻城黑山羊群体中仅有AA和GA 2种基因型外,其余6个SNPs位点在麻城黑山羊、波尔山羊和黑头羊群体中均表现为CC(AA)、CT(GA)和TT(GG)3种基因型。波尔山羊群体中g.9734 A>G、g.9772 T>C位点表现为低度多态(PIC<0.25);麻城黑山羊群体中g.9734 G>A和g.7919 G>A位点表现为低度多态(PIC<0.25),g.9772 T>C、g.352 C>T、g.955 C>T、g.1880 G>A和g.2566 T>C位点表现为中度多态(0.25G、g.352 C>T和g.7919 G>A位点表现为低度多态(PIC<0.25)。卡方适应性检测结果表明,g.9734 A>G、g.9772 T>C和g.352 C>T位点在3个山羊群体中均处于Hardy-Weinberg平衡状态(P>0.05),波尔山羊及麻城黑山羊群体中g.2566 T>C位点处于Hardy-Weinberg平衡状态(P>0.05)。关联分析结果显示,g.9772 T>C位点与波尔山羊头胎产羔数显著关联(P<0.05),g.352 C>T和g.7919 G>A位点与波尔山羊总体产羔数均显著或极显著关联(P<0.05;P<0.01);g.352 C>T位点与黑头羊总体产羔数极显著关联(P<0.01);g.9772 T>C位点与麻城黑山羊头胎产羔数显著关联,g.9734 A>G和g.9772 T>C位点与麻城黑山羊总体产羔数均显著关联(P<0.05)。连锁不平衡分析结果表明,波尔山羊群体中g.352 C>T、g.2566 T>C、g.7919 G>A和g.9734 A>T位点之间处于强连锁平衡状态(D’>0.85,r2=1);黑头羊群体中g.352 C>T和g.2566 T>C位点之间及g.2566 T>C、g.9734 A>T和g.9772 T>C位点之间处于强连锁平衡状态(D’>0.83,r2=1);麻城黑山羊群体中g.352 C>T和g.9772 T>C位点之间处于强连锁平衡状态(D’>0.87,r2=1)。【结论】本研究筛选出7个SNPs,在山羊群体中共存在5个完全连锁的SNPs,其中g.352 C>T、g.7919 G>A、g.9772 T>C和g.9734 A>G 4个SNPs位点可分别作为特定山羊品种育种过程中产羔性能的潜在候选遗传标记,为山羊产羔性能研究提供参考数据。

【Objective】 The purpose of this experiment was to study the association between single nucleotide polymorphism(SNP) in 3′-UTR,intron 1 and exon 7 of Z-DNA binding protein 1(ZBP1) gene and lambing performance in goats, and provide new genetic markers for molecular breeding of high fertility goats.【Method】 Macheng Black goats, Black headed goats and Boer goats were selected as research samples, the blood samples were collected and DNA was extracted.Primers were designed according to the location information of SNP selected from transcriptome data in ZBP1 gene sequence of goats.Multiple single base extension SNP typing technique(SNaPshot) was used to analyze the genetic diversity of ZBP1 gene SNP.General linear model(GLM) was used to analyze the relationship between SNP of ZBP1 gene and lambing performance of 3 goat breeds.【Result】 There were 12 potential SNPs of ZBP1 gene in goats, of which 7 SNPs(g.9734 A>G,g.9772 T>C,g.352 C>T,g.955 C>T,g.1880 G>A,g.2566 T>C and g.7919 G>A) were polymorphic.There were 2 genotypes of AA and GA at g.9734 A>G in Macheng Black goats population, the other 6 SNPs showed 3 genotypes including CC(AA),CT(GA) and TT(GG) in Macheng Black goats, Boer goats and Black headed goats.g.9734 A>G and g.9772 T>C showed low polymorphism in Boer goats population(PIC<0.25),g.9734 G>A and g.7919 G>A showed low polymorphism in Macheng Black goats population(PIC<0.25),while g.9772 T>C,g.352 C>T,g.955 C>T,g.1880 G>A and g.2566 T>C showed moderate polymorphism in Black headed goats population(0.25G,g.352 C>T and g.7919 G>A showed low polymorphism in Black headed goats(PIC<0.25).The chi-square fitness test results show that g.9734 A>G,g.9772 T>C and g.352 C>T were in Hardy-Weinberg equilibrium in 3 goat populations(P>0.05),g.2566 T>C in Boer goats and Macheng Black goats population was in Hardy-Weinberg equilibrium(P>0.05).Correlation analysis results showed that g.9772 T>C was significantly associated with the first litter sizes in Boer goats(P<0.05),g.352 C>T and g.7919 G>A were significantly or extremely significantly associated with the overall litter sizes in Boer goats(P<0.05;P<0.01).g.352 C>T were extremely significantly associated with the overall litter sizes in Black head goat(P<0.01).g.9772 T>C was significantly associated with the first litter sizes in Macheng Black goats, g.9734 A>G and g.9772 T>C were significantly associated with the overall litter sizes in Macheng Black goats(P<0.05).Linkage disequilibrium(LD) results showed that there were strong linkage disequilibrium between g.352 C>T,g.2566 T>C,g.7919 T>C and g.9734 A>T in Boer goats,(D’>0.85,r~2=1). g.352 C>T and g 2566 T>C,and g.2566 T>C,g.9734 A>T and g.9772 T>C in Black headed goats were in strong linkage equilibrium(D’>0.83,r~2=1).There were strong linkage disequilibrium between g.352 C>T and g.9772 T>C in Macheng Black goats(D’>0.87,r~2=1).【Conclusion】 There were 7 SNPs were found in this study, 5 fully linked SNPs were idetified on ZBP1 gene, which indicated that the 4 SNPs(g.352 C>T,g.7919 G>A,g.9772 T>C and g.9734 A>G) could be used as potential candidate genetic markers for lambing performance in the breeding process of specific goat breeds, it provided reference data for molecular marker saaisted breeding of goats.