本文選題：基因組　切入點(diǎn)：重測(cè)序　出處：《西北農(nóng)林科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：部分綿羊具有四個(gè)或更多的乳頭,但有關(guān)這一性狀的遺傳機(jī)理研究報(bào)道較少。為了探究其遺傳差異,本研究對(duì)四乳頭湖羊基因組進(jìn)行了重測(cè)序分析,篩選出與該性狀相關(guān)的候選基因。1、采集四乳頭湖羊(試驗(yàn)組)和兩乳頭湖羊(對(duì)照組)血液樣本,提取血樣DNA進(jìn)行全基因組重測(cè)序,測(cè)序結(jié)果與綿羊參考基因組(Ovis aries Oar_3.1)比對(duì),結(jié)果表明:(1)使用GATK算法比對(duì)測(cè)序結(jié)果,查找湖羊基因組的突變位點(diǎn)。其中在基因編碼區(qū)中,試驗(yàn)組中共有42001個(gè)SNP/InDel位點(diǎn),對(duì)照組共有50010個(gè)SNP/InDel位點(diǎn)。(2)對(duì)SNP/InDel位點(diǎn)進(jìn)行維恩分析,得到試驗(yàn)組湖羊特有的SNP/InDel位點(diǎn)39個(gè),提示這些突變位點(diǎn)可能是四乳頭性狀的潛在關(guān)聯(lián)位點(diǎn)。(3)對(duì)試驗(yàn)組特有的39個(gè)SNP/InDel位點(diǎn)進(jìn)行注釋,共計(jì)注釋到21個(gè)基因。并對(duì)這21個(gè)基因進(jìn)行GO富集分析和KEGG通路分析,涉及物質(zhì)代謝、免疫應(yīng)答、細(xì)胞生長(zhǎng)、機(jī)體發(fā)育和信號(hào)通路等功能。2、對(duì)湖羊基因組重測(cè)序得到試驗(yàn)組中的33個(gè)SNP位點(diǎn)進(jìn)行了驗(yàn)證,利用Sanger測(cè)序法和SNaPshot技術(shù)對(duì)變異位點(diǎn)進(jìn)行SNP分型,結(jié)果表明:(1)與參考基因組對(duì)比,33個(gè)變異位點(diǎn)中,有18個(gè)位點(diǎn)堿基沒有發(fā)生改變,15個(gè)位點(diǎn)發(fā)生了堿基變化。其中ZEB1基因中sheep13-32879407位點(diǎn)和DEPDC7基因中sheep15-61871375位點(diǎn)試驗(yàn)組相比于對(duì)照組存在較高的突變率。sheep13-32879407GC位點(diǎn)在四乳頭樣本中的突變率為91%,在對(duì)照組中的突變率為30%;sheep15-61871375GC位點(diǎn)在四乳頭樣本中的突變率為72%,在對(duì)照組中的突變率為29%。(2)對(duì)ZEB1基因的sheep13-32879407GC位點(diǎn)與湖羊四乳頭發(fā)育等級(jí)進(jìn)行關(guān)聯(lián)分析。在總樣本中,基因型CC和GC分別比基因型GG個(gè)體擁有更高的四乳頭等級(jí)(P0.01)。在試驗(yàn)組中,基因型CC和GC分別比基因型GG個(gè)體擁有更高的四乳頭等級(jí)(P0.05)。說明這兩個(gè)基因型的個(gè)體四乳頭發(fā)育更好。對(duì)DEPDC7基因的sheep15-61871375AC位點(diǎn)與湖羊四乳頭等級(jí)進(jìn)行關(guān)聯(lián)分析。在總樣本中,基因型CC和AC分別比基因型GG個(gè)體擁有更高的四乳頭等級(jí)(P0.01),說明這兩個(gè)基因型個(gè)體的四乳頭發(fā)育更好。綜上所述,ZEB1和DEPDC7基因可作為四乳頭性狀的候選基因。
[Abstract]:Some sheep have four or more nipples, but there are few studies on the genetic mechanism of this trait. The candidate gene. 1. Blood samples were collected from four-tipped Hu sheep (test group) and two nipple lake sheep (control group). The whole genome was sequenced by DNA. The results were compared with that of sheep reference genome Ovis aries Oar3.1). The results showed that GATK algorithm was used to compare the results of sequencing and to find mutation sites in Hu sheep genome. In the gene coding region, there were 4 200 SNP/InDel loci in the experimental group and 50010 SNP/InDel loci in the control group. 39 SNP/InDel loci specific to Hu sheep were obtained in the experiment group, indicating that these mutation sites may be the potential association sites for tetra nipple traits.) the 39 SNP/InDel loci of the test group were annotated. A total of 21 genes were annotated. The 21 genes were analyzed by go enrichment analysis and KEGG pathway analysis, involving substance metabolism, immune response and cell growth. In this study, 33 SNP loci in the experimental group were confirmed by re-sequencing of the Hu sheep genome. Sanger sequencing and SNaPshot techniques were used to type the mutation loci. The results showed that: 1) compared with the reference genome, 33 mutation loci, The mutation rate of sheep13-32879407 locus in ZEB1 gene and sheep15-61871375 locus in DEPDC7 gene was higher than that in control group. The mutation rate was 91. The mutation rate in the control group was 30 sheep15-61871375GC in the four nipple samples, and the mutation rate in the control group was 29.2. The relationship between the sheep13-32879407GC locus of the ZEB1 gene and the development grade of the fourth nipple of Hu sheep was analyzed. Genotype CC and GC had higher levels of four nipples (P0.01) than genotype GG. In the trial group, the four nipple levels of CC and GC were higher than those of genotype GG. The genotype CC and GC had higher four nipple grade (P0.05) than those of genotype GG, which indicated that the four nipples of the two genotypes had better development. The sheep15-61871375AC loci of the DEPDC7 gene were correlated with the four nipple grades of Hu sheep. The genotype CC and AC had higher four nipple grade (P0.01) than those of genotype GG, which indicated that the four nipples of the two genotypes had better development. In conclusion, ZEB1 and DEPDC7 genes could be used as candidate genes for the tetra nipple traits.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S826

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