三个家马品种泌乳相关基因多态性及其与伊犁马产奶量的关联研究

发布时间：2018-04-18 14:13

本文选题：家马 + 产奶量　；参考：《西北农林科技大学》2017年硕士论文

【摘要】：马奶含有丰富的维生素和矿物质,营养价值极高,但其产量远低于牛奶。随着人民生活水平的提高,马奶的需求量增加,加强马泌乳遗传基础研究,开发相关的分子标记,培育乳用马品种势在必行。本研究选择了9个家畜泌乳性能候选基因:催乳素(PRL)及催乳素受体基因(PRLR)、垂体特异性转录因子1(POU1F1)、生长激素1(GH1)及生长激素受体基因(GHR)、乳铁蛋白基因(LTF)、雌激素受体基因(ESR1、ESR2)和三磷酸腺苷结合转运蛋白G超家族成员2基因(ABCG2),根据NCBI提供的基因序列设计扩增CDS区序列,以3个中国家马品种(伊犁马、关中马和蒙古马)共337匹马为研究对象,测序检测鉴定候选基因CDS区的单核苷酸多态位点,采用PCR-RFLP技术对其进行分型,并将伊犁马上发现SNP位点与伊犁马日均产奶量进行单因素方差分析,获得如下研究结果:1.在3个中国家马品种9个泌乳相关基因中检测到10个SNPs(PRL c.210 CT,PRL g.9115 TG,PRL g.9119 CA,PRL g.11090 AG,PRLR g.145227 TG,POU1F1g.2758 TC,GH1 c.22 AC,GH1 g.1584 CG,GHR g.-562 CT,LTF c.172 GA),均为首次发现。其中PRLR g.145227 TG,GHR g.-562 CT,LTF c.172 GA 3个位点仅发现于伊犁马上,PRL g.9115 TG,PRL g.11090 AG,GH1 c.22 AC,GH1 g.1584 CG4个位点仅发现于关中马上,PRL g.9119 CA位点为关中马、蒙古马共有,PRL c.210CT,POU1F1 g.2758 TC位点在三个品种上均有发现。全部10个SNP多态位点中,有3个位点发生在基因的CDS区域,引起错义突变;1个位点发生在基因的启动子区域,其余6个位点均发生在内含子上。2.使用PCR-RFLP方法对伊犁马上检测到的SNP多态进行分型统计,并与其120天日均产奶量进行关联分析。检测到伊犁马上存在的5个不同基因的SNP位点中,PRL c.210 CT和LTF c.172 GA是发生在CDS区上的错义突变,PRL c.210 CT突变导致PRL基因编码蛋白第70位苯丙氨酸突变为亮氨酸,LTF c.172 GA突变致LTF编码蛋白第57位精氨酸突变为赖氨酸;GHR g.-562 CT发生在GHR基因启动子区域,其余位点均位于内含子上。与日均产奶量的关联分析结果显示,5个不同基因的SNP位点中,除PRL c.210 CT外,各位点不同基因型的日均产奶量差异不显著。PRL c.210 CT位点不同基因型的日均产奶量差异显著,其中CC基因型个体日均产奶量显著高于CT基因型个体,CT基因型个体日均产奶量与TT基因型个体差异不显著,推测C等位基因可能对伊犁马产奶量性状有正遗传效应。3.使用双荧光素酶报告基因系统,检测马PRL、PRLR基因启动子活性区域。检测结果显示,马PRL基因上游-418~-1050bp区间为该基因的主要活性区域,其中-418~-738bp区间存在较强的顺势作用元件;马PRLR基因启动子活性区域位于上游-702~-2399bp区间,-1546~-1798bp区间存在较强的顺势作用元件。目前,中国家马品种泌乳性能相关基因的研究甚少。本研究初步调查了3个中国家马品种9个泌乳相关基因CDS区SNP情况,并将伊犁马上发现SNP位点与其日均产奶量进行关联分析,初步探索了马PRL、PRLR基因启动子活性区域,为中国乳用马的高效选育、种质资源保存与利用提供了一定的依据。
[Abstract]:Milk is rich in vitamins and minerals, high nutritional value, but its yield is much lower than that of milk. With the improvement of people's living standard, demand for milk increase, to strengthen the study of the genetic basis of horse lactation, development of molecular markers related to the breeding of dairy horse breeds is imperative. This study selected 9 livestock lactation performance candidate genes: prolactin (PRL) and prolactin receptor gene (PRLR), pituitary specific transcription factor 1 (POU1F1) 1 (GH1), growth hormone and growth hormone receptor gene (GHR), lactoferrin (LTF) gene, estrogen receptor gene (ESR1, ESR2) and ATP binding cassette superfamily G members of the 2 gene (ABCG2), according to the amplification of sequences of CDS gene sequence design provided by NCBI, with 3 China home horse breeds (Yili horse, Mongolia horse and horse Guanzhong) a total of 337 horses as the research object, the detection and identification of candidate gene CDS region nucleotide sequencing Polymorphic loci, carries on the classification using PCR-RFLP technology, and Yili will immediately find SNP site and Yili Marie are milk production by single factor variance analysis, the main results are as follows: 10 to 1. in 9 lactation SNPs detected 3 genes related to Chinese horse breeds (PRL c.210 CT, PRL g.9115 TG PRL, g.9119 CA, PRL g.11090 AG, PRLR g.145227 TG, POU1F1g.2758 TC, GH1 c.22 AC, GH1 g.1584 CG, GHR g.-562 CT, LTF c.172, GA) were found for the first time. The PRLR g.145227 TG, GHR g.-562 CT, LTF c.172 GA 3 loci was found immediately PRL g.9115 TG Li huy, PRL. G.11090 AG, GH1 c.22 AC, GH1 g.1584 CG4 loci were found only in the PRL g.9119 CA site immediately, for a total of Guanzhong horse, Mongolia horse, PRL c.210CT, POU1F1 g.2758 TC loci were found in three cultivars. All 10 SNP loci, 3 loci in CDS gene Area caused by missense mutations; 1 loci in the promoter regions of genes, the remaining 6 loci occurred in the intron of.2. using the PCR-RFLP method to detect the SNP polymorphism in Yili immediately classified statistics, and the 120 day average milk production for the correlation analysis. SNP loci of 5 different genes detected Yili immediately exists in the PRL c.210 CT and LTF c.172 GA in CDS region is a missense mutation, PRL c.210 CT mutation in the PRL gene encoding protein seventieth mutated to phenylalanine leucine, LTF c.172 GA induced LTF mutation protein encoding fifty-seventh mutation of arginine to lysine; GHR g.-562 CT start the sub region in the GHR gene, the other sites were located in introns. The correlation analysis result and average daily milk yield showed that 5 different gene loci of SNP, c.210 in addition to PRL CT, average daily milk yield differences between different genotypes have not The average daily milk yield were.PRL c.210 CT loci had significant difference, the CC genotype average daily milk yield was significantly higher than that of the CT genotype, CT genotype average daily milk yield and TT genotypes were not significantly different, suggesting that C allele may have positive genetic effect of.3. using dual luciferase gene system on Yili horse milk production traits, detection of equine PRL, PRLR gene promoter region. Test results show that the horse PRL gene upstream -418~-1050bp interval as the main active region of the -418~-738bp gene, which has a strong range of cis acting element; Ma PRLR gene promoter region located upstream of the -702~-2399bp interval, -1546~-1798bp interval exists homeopathic components strong. At present, the research Chinese home lactation performance of horse breeds related genes understood. We investigated the 3 Chinese horse breed 9 mi The SNP status in the CDS region of the milk related gene was analyzed, and the correlation between the SNP loci and the daily milk yield was discovered in Yili. The active region of horse PRL and PRLR gene promoter was preliminarily explored, which provided a basis for the efficient breeding of Chinese milk horse and the preservation and utilization of germplasm resources.

【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S821

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