本文關(guān)鍵詞： GPAM基因 單核苷酸多態(tài)性 過表達(dá)載體 干擾載體 奶牛乳腺上皮細(xì)胞 甘油三酯　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：甘油-3-磷酸�；D(zhuǎn)移酶(GPAM)是合成甘油三酯的第一步反應(yīng)的關(guān)鍵酶,對(duì)乳脂形成過程具有很大的影響。本實(shí)驗(yàn)旨在研究GPAM基因?qū)χ袊?guó)荷斯坦奶牛乳脂水平的影響。在本研究中,先采用PCR-PFLP的方法對(duì)GPAM基因遺傳多態(tài)性進(jìn)行了檢測(cè),進(jìn)一步分析GPAM基因SNP位點(diǎn)與中國(guó)荷斯坦奶牛產(chǎn)奶性狀的相關(guān)性。實(shí)驗(yàn)結(jié)果表明:在中國(guó)荷斯坦奶牛群體中,檢測(cè)到了三個(gè)SNP位點(diǎn),分別為E20-3386GA位點(diǎn)、I18-652AG位點(diǎn)和I18-726AG位點(diǎn)。其中,牛GPAM基因E20-3386GA位點(diǎn)與中國(guó)荷斯坦奶牛乳蛋白率及體細(xì)胞數(shù)呈顯著相關(guān)性(p0.05);I18-652AG位點(diǎn)與中國(guó)荷斯坦奶牛乳蛋白、干物質(zhì)及體細(xì)胞數(shù)呈顯著相關(guān)性(p0.05);I18-726AG與中國(guó)荷斯坦奶牛乳蛋白率、奶量、干物質(zhì)、體細(xì)胞數(shù)均呈顯著相關(guān)性(p0.05)。其次構(gòu)建了牛GPAM基因過表達(dá)載體及干擾載體,轉(zhuǎn)染奶牛乳腺上皮細(xì)胞后,對(duì)GPAM基因與脂代謝的相關(guān)功能進(jìn)行驗(yàn)證。實(shí)驗(yàn)結(jié)果表明:在奶牛乳腺上皮細(xì)胞中,牛GPAM基因能調(diào)節(jié)脂肪代謝信號(hào)通路中關(guān)鍵因子AGPAT4基因的m RNA表達(dá)水平,當(dāng)GPAM基因過表達(dá)時(shí),AGPAT4基因的m RNA表達(dá)水平顯著上升;當(dāng)GPAM基因沉默時(shí),AGPAT4基因的m RNA和蛋白表達(dá)水平顯著下降。除此之外,牛GPAM基因過表達(dá)時(shí),奶牛乳腺上皮細(xì)胞中的甘油三酯含量顯著上升;牛GPAM基因沉默時(shí),奶牛乳腺上皮細(xì)胞中的甘油三酯含量顯著下降。實(shí)驗(yàn)結(jié)果表明:GPAM基因可能在調(diào)節(jié)動(dòng)物脂肪代謝中起關(guān)鍵作用。
[Abstract]:Glycerol-3-phosphate transferase (GPAM) is the key enzyme in the first step of synthesis of triglycerides. The aim of this study was to study the effect of GPAM gene on milk fat level in Chinese Holstein cows. The genetic polymorphism of GPAM gene was detected by PCR-PFLP. The relationship between the SNP locus of GPAM gene and milk production traits of Chinese Holstein cows was further analyzed. The results showed that three SNP loci were detected in Chinese Holstein dairy cattle population. They are E20-3386GA locus I18-652AG and I18-726AG, respectively. The E20-3386GA locus of bovine GPAM gene was significantly correlated with milk protein rate and somatic cell number of Chinese Holstein cows. The I18-652AG locus was significantly correlated with milk protein, dry matter and somatic cell number of Chinese Holstein cows. I18-726 AG and Chinese Holstein Dairy cows milk protein rate, milk quantity and dry matter. Bovine GPAM gene overexpression vector and interference vector were constructed after transfection of bovine mammary epithelial cells. The function of GPAM gene and lipid metabolism was verified. The results showed that: in dairy cow mammary epithelial cells. Bovine GPAM gene can regulate the expression level of m RNA of AGPAT4 gene, a key factor in lipid metabolism signaling pathway, when GPAM gene is overexpressed. The expression level of m RNA of AGPAT4 gene increased significantly. When the GPAM gene was silenced, the expression level of m RNA and protein of AGPAT4 gene decreased significantly. In addition, the overexpression of bovine GPAM gene was observed. The content of triglyceride in dairy cow mammary epithelial cells increased significantly. The content of triglyceride in bovine mammary epithelial cells decreased significantly when the bovine GPAM gene was silenced. The results showed that the GPAM gene may play a key role in regulating animal fat metabolism.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S823

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1 趙堯璐;GPAM基因遺傳多態(tài)性與奶牛乳脂性狀的相關(guān)分析及功能驗(yàn)證[D];吉林大學(xué);2017年

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本文編號(hào)：1467009