【摘要】：隨著基因組學(xué)研究以及基因芯片技術(shù)的發(fā)展,人們通過全基因組關(guān)聯(lián)分析(GWAS)的研究策略發(fā)現(xiàn)了大量影響復(fù)雜性狀的遺傳變異。近年來,該方法在篩選和鑒定農(nóng)業(yè)動物重要經(jīng)濟性狀的主效基因和因果位點中得到了廣泛應(yīng)用。豬作為國內(nèi)最重要的肉類供應(yīng)動物,對其重要經(jīng)濟性狀的研究更受關(guān)注。本研究以五個實驗群體:F2資源家系群體、蘇太群體、萊蕪群體、二花臉群體和杜長大商業(yè)群體共2650個體為研究對象,利用豬60K SNP芯片對這些個體進行高通量基因分型,并記錄內(nèi)臟(心臟、肝臟、脾臟和腎臟)重和胴體重以及C20:0脂肪酸含量等重要經(jīng)濟性狀,采用GWAS及Meta-GWAS的策略進行分析,擬定位與這些性狀顯著相關(guān)的主效位點。在此基礎(chǔ)上,綜合運用單倍型分析、IBD分析、斷點重組分析等多種統(tǒng)計學(xué)手段精細定位影響C20:0含量相關(guān)的數(shù)量性狀位點(QTL),旨在找到影響C20:0的數(shù)量性狀基因(QTG)以及找到相應(yīng)的數(shù)量性狀核苷酸(QTN),進一步深入解析豬16號染色體上影響肌肉中C20:0的主效QTL的分子遺傳機理。結(jié)果表明:1)利用豬60K芯片在五個群體中共檢測到10個基因組顯著水平的QTL,4個與心臟重相關(guān),2個與肝臟重相關(guān),2個與脾臟重相關(guān),1個與胴體重相關(guān),1個與C20:0相關(guān)。此外,本研究還定為到一個一因多效的QTL,7號染色體上的QTL同時影響心臟重和肝臟重。2)采用meta-GWAS分析后,有62%的達到基因組顯著水平的QTL在Meta分析中得到了驗證,并且新現(xiàn)了4個顯著影胴體重的QTL。3)利用單倍型分析、IBD分析以及Meta分析等研究策略,將C20:0含量定位于41.3-43.4 Mb處,并在這一區(qū)間找到了影響C20:0的候選基因ELOVL7。4)利用實驗室6頭F0重測序數(shù)據(jù),挖掘出11個與QTL基因型相符合的SNPs,經(jīng)過在蘇太群體中的驗證,最后剩下(42561841和42564441bp)2個候選SNPs。5)為了驗證ELOVL7對C20:0脂肪酸的生物學(xué)影響,隨機選取120個F2個體的眼肌進行RNA定量表達實驗,利用60K芯片結(jié)果進行eQTL GWAS分析,在SSC16 40.7Mb(位于ELOVL7附近)處檢測到顯著影響C20:0 RNA表達量的標(biāo)記ALGA0090412。本研究將為后期鑒別內(nèi)臟和胴體重的遺傳結(jié)構(gòu)以及深入解析豬肉中C20:0含量變異的遺傳結(jié)構(gòu)奠定基礎(chǔ)。
[Abstract]:With the development of genomics and gene chip technology, a large number of genetic variations affecting complex traits have been discovered through the research strategy of genome-wide association analysis (GWAS). In recent years, this method has been widely used in the screening and identification of major genes and causal loci of important economic traits in agricultural animals. As the most important meat supplier in China, pigs have been paid more attention to their important economic characters. In this study, a total of 2650 individuals from five experimental populations: F _ 2 resource family, Sutai, Laiwu, Erhualian and du Changlong commercial population were selected as subjects. The high throughput genotyping of these individuals was carried out by using 60 K SNP chip. The visceral weight (heart, liver, spleen and kidney), carcass weight and C20: 0 fatty acid content were recorded and analyzed by GWAS and Meta-GWAS. On this basis, the haplotype analysis and IBD analysis are used synthetically. (QTL), a quantitative trait locus related to C20: 0 content, is designed to find the quantitative trait gene (QTG), which affects C20: 0, and to find the nucleotide (QTN), of the corresponding quantitative trait to further analyze the quantitative trait locus, such as breakpoint recombination analysis and so on. Molecular genetic mechanism of dominant QTL in muscle affected by C20: 0 on chromosome 16. The results showed that 10 genome-wide significant levels of QTL, were detected in five populations using 60 K porcine microarray. Four of them were correlated with heart weight, two with liver weight, two with spleen weight, one with carcass weight and one with C20: 0. In addition, this study was determined to be a multieffectual QTL on chromosome 7 of QTL, that affects both heart and liver weights. After meta-GWAS analysis, 62% of QTL with genomic significance were verified by Meta analysis. And four new QTL.3 with significant carcass weight were developed. By using haplotype analysis and Meta analysis, the content of C20: 0 was located at 41.3-43.4 Mb, and the content of C20: 0 was located at 41.3-43.4 Mb, and the content of C20: 0 was located at 41.3-43.4 Mb. The candidate gene ELOVL7.4, which affects C20: 0, was found in this region. Using 6 F0 resequenced data from the laboratory, 11 SNPs, corresponding to the QTL genotype were found to have been verified in the Sutai population. In order to verify the biological effect of ELOVL7 on C20: 0 fatty acids, 120 F2 individuals were randomly selected for RNA quantitative expression experiment. EQTL GWAS analysis was performed using 60K microarray. Marker ALGA0090412. was detected at SSC16 40.7Mb (near ELOVL7) that significantly affected the expression of C20: 0 RNA This study will lay a foundation for the identification of the genetic structure of viscera and carcass weight and the further analysis of the genetic structure of the variation of C20: 0 content in pork.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S828

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