ZP基因的变异及其对湖羊产羔数的遗传效应研究

发布时间：2018-05-27 18:45

本文选题：透明带蛋白 + 产羔数　；参考：《华中农业大学》2015年硕士论文

【摘要】：本研究主要探讨ZP基因家族(ZP2、ZP3、ZP4)中基因的变异对绵羊产羔数的影响,以寻找与高繁殖力相关的SNP位点。以湖羊、寒羊、蒙古羊和滩羊为试验材料,采用混合样测序技术寻找ZP基因变异的SNP位点,搜集整理了100只湖羊的产羔数数据,通过分析SNP位点与产羔数之间的关系,初步筛选出对产羔数有较大影响的SNP位点。采用生物信息学的方法对ZP蛋白的一级、二级及三级结构进行分析,通过分析ZP基因变异位点和ZP蛋白结构、自然选择的关系,来阐述基因结构与基因功能之间的联系,为羊的选种、育种工作提供理论参考。利用NCBI公共数据库已有的ZP基因序列,通过比较哺乳类、鸟类和鱼类的生理性受精方式和进化状况的差异,来探究驱动ZP基因快速进化的机制。研究结果如下:1.采用混合样测序法共检测到28个SNP位点,其中ZP2有4个位置碱基发生突变,ZP3有7个位置碱基发生突变,ZP4有17个碱基发生突变。SNP间的连锁分析发现,ZP2基因的4个SNP位点形成了1个连锁区块,ZP3基因的T678C和A859C、T7781G和G7804A形成了2个连锁区块和3个独立的SNP位点,ZP4基因形成了3连锁区块和4个独立的SNP位点。将连锁的SNP区块、独立的SNP位点分别与湖羊第一胎产羔数、胎均产羔数进行关联分析后发现,ZP3基因T8176C位点对湖羊的胎产羔数有显著影响,ZP4基因有3个SNP位点(A7499C、T10223C、G10383A)对胎产羔数有明显的影响。结构分析表明,T8176C位于ZP3的跨膜结构域上,ZP4基因的G10383A位于ZP4的ZP结构域上。群体间进化检验发现,ZP3和ZP4基因在绵羊群体受到了正选择的作用。2.对绵羊ZP基因的生物信息学预测显示,绵羊ZP2基因编码713个氨基酸,编码产物是一个偏碱性的亲水跨膜蛋白。信号肽位于氨基酸序列的1-36区域,跨膜结构位于685-704区段,1-684区段位于膜的外部,705-713区段位于膜的内部。二级结构预测显示螺旋占4.63%,延伸链占39.97%,环状结构占55.40%。保守域分析显示ZP2蛋白N端和C端是变异区,中间部分约400个氨基酸较为保守,ZP结构域位于368-629区段。绵羊ZP3基因编码381个氨基酸,编码产物是一个无信号肽的亲水跨膜蛋白。二级结构主要以环状结构为主,其中螺旋占3.94%,延伸链占35.96%,环状结构占60.1%。3-261区段是其ZP结构域,跨膜结构域在341-363的α螺旋区域,在ZP3的239-340区段有一特殊结构,该结构可能是ZP3精卵结合域,305-308位点可能是弗林蛋白酶水解位点。在膜内的371位点的丝氨酸(Ser)受到了蛋白质激酶(PKA)的磷酸化作用,可能具有信号转导的功能。绵羊ZP4基因编码534个氨基酸,编码产物是含有信号肽的亲水跨膜蛋白。二级结构中α-螺旋占4.68%,β-折叠占32.02%,无规则卷曲占63.30%。信号肽位于1-19氨基酸序列,保守结构域分析表明三叶草结构(Trefoil)位于141-183区段,ZP结构域位于188-460区段,跨膜区位于508-530区段。三叶草结构上有2个精卵特异性结合位点和精卵特异性结合环状结构。3.通过分析比对哺乳类、鸟类及鱼类ZP基因的进化情况发现,哺乳类和鱼类的ZP基因具有较快的变异率,受到更大的选择压,而鸟类的变异较慢。正选择分析发现单精入卵类的物种(哺乳动物和鱼类)的ZP基因都经历过快速的进化,而鸟类的5个ZP基因均不受正选择的作用。结构分析显示,大部分的正选择位点分布在ZP2的NTR区域和ZP3的SE区域。
[Abstract]:In this study, the effect of genetic variation in the ZP gene family (ZP2, ZP3, ZP4) on the number of lambs in sheep was investigated in order to find the SNP loci related to high fecundity. In order to find the SNP loci of the ZP gene mutation with the mixed sample sequencing technology, the mixed sample sequencing technology was used to find the SNP loci of the mutation of the ZP gene. The data of the number of lambing of 100 sheep were collected and collected. The relationship between the SNP site and the number of lambs was analyzed, and the SNP loci which had a greater impact on the number of lambs were screened. Bioinformatics was used to analyze the first, two and three grade structure of ZP protein. The relationship between the ZP gene mutation site and the structure of the ZP protein was analyzed to explain the genetic structure and gene function. The relationship between sheep breeding and breeding is a theoretical reference. Using the ZP gene sequence existing in the NCBI public database, the mechanism of the rapid evolution of ZP genes is explored by comparing the differences in the physiological fertilization and evolution of mammals, birds and fishes, and the results are as follows: 1. a total of 28 was detected by the mixed sample sequencing method. SNP loci, in which ZP2 has 4 bases mutation, ZP3 has 7 bases mutation, and ZP4 has 17 base mutation.SNP linkage analysis found that 4 SNP loci of the ZP2 gene form 1 chain blocks, ZP3 gene T678C and A859C, T7781G and G7804A form 2 linkage blocks and 3 independent locus of restriction 3 chain blocks and 4 independent SNP loci were formed. After the linkage analysis of SNP blocks, independent SNP loci with the number of first births and the number of fetal lambs, it was found that the T8176C locus of the ZP3 gene had a significant influence on the number of fetal lambs in the sheep, and the ZP4 gene had 3 SNP loci (A7499C, T10223C, G10383A) for the number of fetal lambs. The structural analysis showed that T8176C was located in the transmembrane domain of ZP3, and the G10383A of ZP4 gene was located on the ZP domain of ZP4. The interpopulation evolution test found that the ZP3 and ZP4 genes were selected in the sheep population by the positive effect of.2. on the bioinformatics of the sheep ZP gene, and the ZP2 gene of sheep encodes 713 amino acids and encoded products. It is an alkaline hydrophilic transmembrane protein. The signal peptide is located in the 1-36 region of the amino acid sequence, the transmembrane structure is located in the 685-704 section, the 1-684 section is located outside the membrane, and the 705-713 section is located inside the membrane. The two stage structure prediction shows that the spiral accounts for 4.63%, the extension chain is 39.97%, the ring structure accounts for the 55.40%. conservative domain analysis to show the N and C ends of the ZP2 protein. About 400 amino acids in the middle part are more conservative and the ZP domain is located in the 368-629 section. The sheep ZP3 gene encodes 381 amino acids, and the encoding product is a hydrophilic transmembrane protein without signal peptide. The two structure is mainly ring structure, in which the spiral accounts for 3.94%, the extension chain is 35.96%, and the ring structure accounts for the 60.1%.3-261 section of its ZP The domain, the trans membrane domain in 341-363 alpha helix region, has a special structure in the 239-340 section of ZP3, which may be the ZP3 sperm binding domain and the 305-308 site may be the Flynn protease site. The serine (Ser) at the 371 site in the membrane is phosphorylated by the protein kinase (PKA) and may have signal transduction. Yes. The sheep ZP4 gene encodes 534 amino acids, the encoding product is a hydrophilic transmembrane protein containing signal peptide. In the two grade structure, the alpha helix accounts for 4.68%, the beta folding accounts for 32.02%, the irregular curl accounts for the 1-19 amino acid sequence of the 63.30%. signal peptide, and the conservative domain analysis shows that the Trefoil is located in the 141-183 section, and the ZP domain is located in 188-46. In the 0 section, the transmembrane region is located in the 508-530 section. There are 2 sperm specific binding sites and sperm specific binding ring structure.3. on the structure of clover. By analyzing the evolution of the ZP gene of mammals, birds and fish, it is found that the ZP gene of mammalian and fishes has a faster variation rate and is subjected to greater selection pressure and the variation of birds. The ZP gene of the species (mammals and fish) of the species (mammals and fish) has undergone rapid evolution, while 5 ZP genes in birds are not affected by positive selection. The structural analysis shows that most of the positive loci are distributed in the NTR region of ZP2 and the SE region of ZP3.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826

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