本文選題：透明帶蛋白 + 產(chǎn)羔數(shù)��； 參考：《華中農(nóng)業(yè)大學》2015年碩士論文

【摘要】：本研究主要探討ZP基因家族(ZP2、ZP3、ZP4)中基因的變異對綿羊產(chǎn)羔數(shù)的影響,以尋找與高繁殖力相關的SNP位點。以湖羊、寒羊、蒙古羊和灘羊為試驗材料,采用混合樣測序技術尋找ZP基因變異的SNP位點,搜集整理了100只湖羊的產(chǎn)羔數(shù)數(shù)據(jù),通過分析SNP位點與產(chǎn)羔數(shù)之間的關系,初步篩選出對產(chǎn)羔數(shù)有較大影響的SNP位點。采用生物信息學的方法對ZP蛋白的一級、二級及三級結構進行分析,通過分析ZP基因變異位點和ZP蛋白結構、自然選擇的關系,來闡述基因結構與基因功能之間的聯(lián)系,為羊的選種、育種工作提供理論參考。利用NCBI公共數(shù)據(jù)庫已有的ZP基因序列,通過比較哺乳類、鳥類和魚類的生理性受精方式和進化狀況的差異,來探究驅動ZP基因快速進化的機制。研究結果如下:1.采用混合樣測序法共檢測到28個SNP位點,其中ZP2有4個位置堿基發(fā)生突變,ZP3有7個位置堿基發(fā)生突變,ZP4有17個堿基發(fā)生突變。SNP間的連鎖分析發(fā)現(xiàn),ZP2基因的4個SNP位點形成了1個連鎖區(qū)塊,ZP3基因的T678C和A859C、T7781G和G7804A形成了2個連鎖區(qū)塊和3個獨立的SNP位點,ZP4基因形成了3連鎖區(qū)塊和4個獨立的SNP位點。將連鎖的SNP區(qū)塊、獨立的SNP位點分別與湖羊第一胎產(chǎn)羔數(shù)、胎均產(chǎn)羔數(shù)進行關聯(lián)分析后發(fā)現(xiàn),ZP3基因T8176C位點對湖羊的胎產(chǎn)羔數(shù)有顯著影響,ZP4基因有3個SNP位點(A7499C、T10223C、G10383A)對胎產(chǎn)羔數(shù)有明顯的影響。結構分析表明,T8176C位于ZP3的跨膜結構域上,ZP4基因的G10383A位于ZP4的ZP結構域上。群體間進化檢驗發(fā)現(xiàn),ZP3和ZP4基因在綿羊群體受到了正選擇的作用。2.對綿羊ZP基因的生物信息學預測顯示,綿羊ZP2基因編碼713個氨基酸,編碼產(chǎn)物是一個偏堿性的親水跨膜蛋白。信號肽位于氨基酸序列的1-36區(qū)域,跨膜結構位于685-704區(qū)段,1-684區(qū)段位于膜的外部,705-713區(qū)段位于膜的內(nèi)部。二級結構預測顯示螺旋占4.63%,延伸鏈占39.97%,環(huán)狀結構占55.40%。保守域分析顯示ZP2蛋白N端和C端是變異區(qū),中間部分約400個氨基酸較為保守,ZP結構域位于368-629區(qū)段。綿羊ZP3基因編碼381個氨基酸,編碼產(chǎn)物是一個無信號肽的親水跨膜蛋白。二級結構主要以環(huán)狀結構為主,其中螺旋占3.94%,延伸鏈占35.96%,環(huán)狀結構占60.1%。3-261區(qū)段是其ZP結構域,跨膜結構域在341-363的α螺旋區(qū)域,在ZP3的239-340區(qū)段有一特殊結構,該結構可能是ZP3精卵結合域,305-308位點可能是弗林蛋白酶水解位點。在膜內(nèi)的371位點的絲氨酸(Ser)受到了蛋白質(zhì)激酶(PKA)的磷酸化作用,可能具有信號轉導的功能。綿羊ZP4基因編碼534個氨基酸,編碼產(chǎn)物是含有信號肽的親水跨膜蛋白。二級結構中α-螺旋占4.68%,β-折疊占32.02%,無規(guī)則卷曲占63.30%。信號肽位于1-19氨基酸序列,保守結構域分析表明三葉草結構(Trefoil)位于141-183區(qū)段,ZP結構域位于188-460區(qū)段,跨膜區(qū)位于508-530區(qū)段。三葉草結構上有2個精卵特異性結合位點和精卵特異性結合環(huán)狀結構。3.通過分析比對哺乳類、鳥類及魚類ZP基因的進化情況發(fā)現(xiàn),哺乳類和魚類的ZP基因具有較快的變異率,受到更大的選擇壓,而鳥類的變異較慢。正選擇分析發(fā)現(xiàn)單精入卵類的物種(哺乳動物和魚類)的ZP基因都經(jīng)歷過快速的進化,而鳥類的5個ZP基因均不受正選擇的作用。結構分析顯示,大部分的正選擇位點分布在ZP2的NTR區(qū)域和ZP3的SE區(qū)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.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S826

【共引文獻】

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1 何余ng;利用全基因組高通量SNP標記定位豬乳頭數(shù)和斷奶體重QTL[D];江西農(nóng)業(yè)大學;2011年

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1 楊志軍;清平豬部分種質(zhì)特性的研究[D];華中農(nóng)業(yè)大學;2008年

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