本文選題：豬 + 選擇信號�。� 參考：《華中農(nóng)業(yè)大學》2017年碩士論文

【摘要】：我國作為第一養(yǎng)豬大國,豬肉類產(chǎn)品占我國肉類消費的65%左右。養(yǎng)豬業(yè)是我國畜牧業(yè)發(fā)展的重點,但目前我國養(yǎng)豬經(jīng)濟效益還有待改善。我國地方豬與西方豬之間經(jīng)濟性狀有著較大差異。西方豬由于長期的人工選擇使得豬的經(jīng)濟性能不斷提高,而中國地方豬由于選擇的目的不同且選擇的強度較弱,目前在瘦肉率、生長速度等重要的經(jīng)濟性狀上與西方豬還存在較大的差距。人工選擇的不同是中國地方豬與西方豬的經(jīng)濟性狀產(chǎn)生較大差異的原因。本研究首先利用高通量測序數(shù)據(jù)對中西方豬種的選擇信號差異進行了分析,篩選中西方豬種中受到強烈選擇的基因,結(jié)果顯示IGF1R、Ras GRP1及Ins R基因在中西方豬種中受到強烈選擇,然后對等位基因差異表達情況進行了研究并進行了位點保守性分析,預測出關(guān)鍵的功能調(diào)控位點進行后續(xù)的分子水平研究及群體研究,以期望找到對豬進行品種改良的關(guān)鍵分子標記。本研究得到的主要結(jié)果如下:(1)通過選擇信號分析發(fā)現(xiàn)IGF1R、Ras GRP1及Ins R基因在中西方豬種中受到強烈選擇,并通過等位基因差異表達研究和物種保守性分析找到3個很可能影響基因表達的候選功能位點。(2)凝膠遷移實驗結(jié)果發(fā)現(xiàn)轉(zhuǎn)錄因子YY1能夠結(jié)合IGF1R基因-61K A/G多態(tài)位點,且對等位基因G有更強的結(jié)合能力。雙熒光素酶報告基因系統(tǒng)活性檢測證明該位點能夠促進下游基因表達,且G等位基因發(fā)揮的作用更強。然后通過RNA干擾實驗發(fā)現(xiàn)轉(zhuǎn)錄因子YY1表達降低能夠抑制IGF1R的表達。(3)凝膠遷移實驗結(jié)果顯示轉(zhuǎn)錄因子Myo D能夠結(jié)合IGF1R基因Intron20 G/A多態(tài)位點,且對G等位基因有更強的結(jié)合能力。然后通過熒光定量PCR發(fā)現(xiàn)小鼠骨骼肌衛(wèi)星細胞分化后IGF1R和Myo D的表達量都著極顯著的升高。(4)對IGF1R基因-61K A/G多態(tài)與杜洛克×二花臉F2群體免疫及生長性狀進行關(guān)聯(lián)分析,結(jié)果表明該位點與33日齡嗜酸性粒細胞比率極顯著關(guān)聯(lián)(p0.01),與80日齡體重顯著關(guān)聯(lián)(p0.05),可以作為對豬生長及免疫性狀改良的候選分子標記。(5)對IGF1R基因Intron20 G/A多態(tài)與杜洛克×二花臉F2群體免疫及生長性狀進行關(guān)聯(lián)分析,結(jié)果表明該位點與33日齡嗜酸性粒細胞比率及80日齡體重都顯著關(guān)聯(lián)(p0.05),可以作為對豬生長及免疫性狀改良的候選分子標記。(6)對RasGRP1基因+1K A/G多態(tài)與杜洛克×二花臉F2群體免疫及生長性狀進行關(guān)聯(lián)分析,結(jié)果表明該位點與80日齡體重顯著關(guān)聯(lián)(p0.05),可以作為對豬生長性狀改良的候選分子標記。
[Abstract]:As the largest pig-raising country in China, pork products account for about 65% of our meat consumption.Pig raising industry is the focus of animal husbandry development in China, but the economic benefit of pig raising in China needs to be improved at present.There are great differences in economic characters between local pigs and western pigs in China.Western pigs have improved their economic performance as a result of long-term artificial selection, while local Chinese pigs are currently in lean meat ratio because of their different purposes and weak selection intensity.There is still a big gap between the growth rate and other important economic traits compared with the western pigs.The difference of artificial selection is the reason for the difference of economic characters between Chinese local pigs and western pigs.In this study, high throughput sequencing data were used to analyze the difference of selection signal between Chinese and Western pig breeds. The results showed that the IGF1RN Ras GRP1 and Ins R genes were strongly selected in Chinese and Western pig breeds.Then the allelic differential expression was studied and the locus conserved analysis was carried out to predict the key functional regulatory sites for further molecular level and population studies.We hope to find the key molecular markers for the improvement of pig breeds.The main results obtained in this study are as follows: (1) through the selection signal analysis, it was found that IGF1RN Ras GRP1 and Ins R genes were strongly selected in Chinese and Western pig breeds.Through allelic differential expression study and species conservation analysis, we found three candidate functional sites that may affect gene expression. The results showed that the transcription factor YY1 could bind to the IGF1R gene -61K A / G polymorphic sites, and the results showed that the transcriptional factor YY1 could bind to the IGF1R gene -61K A / G polymorphic loci.And the allele G has stronger binding ability.The detection of the activity of double luciferase reporter gene system showed that this locus could promote downstream gene expression and G allele played a more important role.Then RNA interference assay showed that the reduced expression of transcription factor YY1 could inhibit the expression of IGF1R. The result of gel migration experiment showed that Myo D could bind to Intron20 G / A polymorphic site of IGF1R gene and had stronger binding ability to G allele.Then fluorescence quantitative PCR showed that the expression of IGF1R and Myo D in mouse skeletal muscle satellite cells were significantly increased after differentiation.) the association analysis between IGF1R gene -61K A / G polymorphism and Duroc 脳 Erhualian F2 population immune and growth traits was carried out.The results showed that this locus was significantly associated with the ratio of eosinophils at 33 days old and weight at 80 days old, and could be used as a candidate molecular marker for the improvement of growth and immune traits of pigs. It could be used as a candidate molecular marker for IGF1R gene Intron20 G / A polymorphism and Duroc gene polymorphism.Ke 脳 Erhualian F2 population immunity and growth traits were analyzed by association analysis.The results showed that this locus was significantly associated with the ratio of eosinophils at 33 days old and body weight at 80 days old, and could be used as a candidate molecular marker for the improvement of growth and immune traits of pigs. It could be used as a candidate molecular marker for RasGRP1 gene 1K A / G polymorphism and Duroc 脳 diflorescence.The immune and growth traits of face F2 population were analyzed by association analysis.The results showed that this locus was significantly associated with body weight at 80 days old and could be used as a candidate molecular marker for improving pig growth traits.
【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】：2017
【分類號】：S828

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