本文選題：ZP3 + 綿羊��； 參考：《華中農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：本研究主要是研究ZP3基因的變異與海拔及產(chǎn)羔數(shù)的關(guān)系,探討ZP3基因在物種間的進化以及在綿羊群體間的選擇類型,尋找ZP3基因與高繁殖力相關(guān)的SNP位點,為綿羊的選育工作提供理論參考。選擇9個不同海拔高度的綿羊群體(高原型藏羊、歐拉型藏羊、山谷型藏羊、烏珠穆沁羊、蘇尼特羊、灘羊、廣靈大尾羊、小尾寒羊和湖羊)作為試驗材料,采用混合樣測序技術(shù)檢測綿羊群體內(nèi)ZP3基因的SNP位點,用直接測序法對其進行分型,對這些位點的基因頻率、基因型頻率與海拔進行關(guān)聯(lián)分析;對15個物種的ZP3基因進行系統(tǒng)發(fā)生關(guān)系、基因相似性及物種間進化分析;對9個綿羊群體內(nèi)檢測到的ZP3基因的SNP位點進行群體受選擇狀況分析;收集整理158只湖羊的產(chǎn)羔數(shù)據(jù),分析SNP位點基因型、連鎖區(qū)單倍型及雙倍型與產(chǎn)羔數(shù)的關(guān)系。研究結(jié)果表明:1.ZP3基因在15個物種間的正選擇檢驗表明ZP3受到強烈的正選擇。模型M2a樸素貝葉斯方法(NEB)檢測發(fā)現(xiàn)ZP3基因不存在正選擇位點,經(jīng)驗貝葉斯方法(BEB)檢測發(fā)現(xiàn)ZP3基因存在3個正選擇位點(364V、371S、372T)。模型M8樸素貝葉斯方法(NEB)檢測發(fā)現(xiàn)ZP3基因存在83個正選擇位點,其中有7個后驗概率顯著的正選擇位點(344Q、364V、371S、372T、374A、399T、401L,P0.95);經(jīng)驗貝葉斯方法(BEB)檢測發(fā)現(xiàn)ZP3基因存在8個正選擇位點(344Q、364V、371S、372T、374A、399T、401L、402P)。這些氨基酸正選擇位點大部分位于精卵特異性識別區(qū)域內(nèi)。2.利用混合樣測序法在綿羊ZP3基因上共檢測到20個SNP位點,其中10個位于內(nèi)含子區(qū)域,10個位于外顯子區(qū)域。g.78TC、g.195GA、g.2293CT、g.7576CT是同義突變,g.259AC和g.7610GA是非同義突變。3.檢測9個綿羊群體ZP3基因序列的差異,利用Lositan對綿羊群體ZP3的SNP位點進行種內(nèi)進化狀況的分析,結(jié)果表明平均分化系數(shù)為0.038,說明只有3.8%的的變異是由群體間產(chǎn)生的,97.2%的變異是由各群體內(nèi)產(chǎn)生的。只有g(shù).2046GA和g.7576CT受到了強烈的正選擇作用,其余位點處于中性選擇。4.9個群體ZP3基因SNP位點的等位基因頻率與海拔進行關(guān)聯(lián)結(jié)果表明:g.7202AG的等位基因A與海拔顯著正相關(guān),等位基因G與海拔顯著負(fù)相關(guān);g.9438GC的等位基因G與海拔顯著正相關(guān),等位基因C與海拔顯著負(fù)相關(guān)。基因型頻率與海拔進行關(guān)聯(lián)分析,結(jié)果表明:g.7179GT的GG基因型與海拔顯著正相關(guān),GT基因型與海拔極顯著負(fù)相關(guān);g.7202AG的AA基因型與海拔顯著正相關(guān),AG基因型與海拔極顯著負(fù)相關(guān);g.9438GC的GG基因型與海拔顯著正相關(guān),GC基因型與海拔顯著負(fù)相關(guān)。5.對湖羊群體SNP基因型、3個連鎖區(qū)塊的單倍型及雙倍型分別與湖羊第1和第2胎產(chǎn)羔數(shù)進行關(guān)聯(lián)分析,結(jié)果表明:位點g.328GA與第1胎產(chǎn)羔數(shù)顯著相關(guān)(P0.05),基因型GA比GG的產(chǎn)羔數(shù)高出1.15個;g.7342AG與產(chǎn)羔數(shù)顯著相關(guān)(P0.05),基因型AG比基AA的產(chǎn)羔數(shù)高出0.66個。SNP基因型與第二胎的關(guān)聯(lián)結(jié)果均不顯著(P0.05)。SNP單倍型、雙倍型與第一、二胎的關(guān)聯(lián)結(jié)果均不顯著(P0.05)。
[Abstract]:The main purpose of this study was to study the relationship between the variation of ZP3 gene and altitude and litter size, to explore the evolution of ZP3 gene among species and the selection type among sheep populations, and to search for the SNP site of ZP3 gene associated with high fecundity. To provide a theoretical reference for the breeding of sheep. Nine sheep populations with different elevations (plateau Tibetan sheep, Euler Tibetan sheep, valley Tibetan sheep, Wuzhimuqin sheep, Sunita sheep, Tan sheep, Guangling large tail sheep, small tail Han sheep and Hu sheep) were selected as experimental materials. The SNP loci of ZP3 gene in sheep population were detected by mixed sample sequencing, and the gene frequency, genotype frequency and altitude of these loci were analyzed by direct sequencing. The phylogenetic relationship, genetic similarity and evolution among species of ZP3 genes of 15 species were analyzed, and the population selection status of SNP loci of ZP3 gene detected in 9 sheep populations were analyzed, and the lambing data of 158 Hu sheep were collected and sorted. The relationship between SNP locus genotype, linkage haplotype and litter size was analyzed. The results showed that: 1. The positive selection test of ZP3 gene in 15 species showed that ZP3 was strongly positively selected. Model M2a naive Bayesian method (NEB) found that there were no positive selection sites in ZP3 gene, and three positive selection sites were found in ZP3 gene by empirical Bayesian method. Model M8 naive Bayesian method (neb) showed that there were 83 positive selection sites in ZP3 gene, among which 7 positive selection sites with significant posttest probability were identified as 344QN 364V ~ 371S ~ 372T ~ 374A ~ (374A) ~ (374A) ~ (399T) ~ 401L ~ (0.95L), and ZP3 gene (n = 8) was found to have 8 positive selection loci 344Q ~ (364V) ~ 371T ~ (372T) ~ (374A) ~ (399T) ~ (401T) ~ (402P). Most of these amino acid positive selection sites are located in the sperm-egg specific recognition region. A total of 20 SNP loci were detected in sheep ZP3 gene by mixed sample sequencing, of which 10 were located in intron region and 10 were located in exon region .g.78TCng. 195GAG. 2293CT.7576CT was synonymous mutation. 259AC and g.7610GA were non-synonymous mutations. 3. The differences of ZP3 gene sequences in 9 sheep populations were detected. The SNP loci of ZP3 in sheep populations were analyzed by Lositan. The results showed that the average differentiation coefficient was 0.038, indicating that only 3.8% of the variation was caused by population variation. Only g.2046GA and g.7576CT were strongly positively selected. The allele frequencies of SNP locus of ZP3 gene in the other loci in neutral selection population were correlated with altitude. The results showed that the allele A of 1. 7 g. 7202 AG was significantly positively correlated with altitude. Allele G was significantly negatively correlated with altitude, allele G was positively correlated with altitude, and allele C was negatively correlated with altitude. Genotype frequency was correlated with altitude. The results showed that the GG genotype of G. 7179GT was significantly positively correlated with the altitude and the AA genotype of G. 7202 AG was significantly negatively correlated with the altitude and the GG genotype of G. 9438GC was significantly negative correlated with the altitude. There was a significant negative correlation between GC genotype and altitude. The SNP genotypes, haplotypes and diplotypes of three linkage blocks were correlated with the first and second litter size of Hu sheep, respectively. The results showed that there was a significant correlation between g.328GA locus and litter size in the first trimester. The number of lambs of genotype GA was 1.15 g 路7342 AG higher than that of GG, and the number of lambs of genotype AG was 0.66 higher than that of AA, and the association between genotype AG and second litter was higher than that of GG. The results showed that the haplotypes of P0.05 and SNP were not significant. The results of correlation between diplotypes and first and second births were not significant (P 0.05).
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S826

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