GPAM基因遗传多态性与奶牛乳脂性状的相关分析及功能验证

发布时间：2018-01-27 00:18

本文关键词： GPAM基因单核苷酸多态性过表达载体干扰载体奶牛乳腺上皮细胞甘油三酯　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：甘油-3-磷酸酰基转移酶(GPAM)是合成甘油三酯的第一步反应的关键酶,对乳脂形成过程具有很大的影响。本实验旨在研究GPAM基因对中国荷斯坦奶牛乳脂水平的影响。在本研究中,先采用PCR-PFLP的方法对GPAM基因遗传多态性进行了检测,进一步分析GPAM基因SNP位点与中国荷斯坦奶牛产奶性状的相关性。实验结果表明:在中国荷斯坦奶牛群体中,检测到了三个SNP位点,分别为E20-3386GA位点、I18-652AG位点和I18-726AG位点。其中,牛GPAM基因E20-3386GA位点与中国荷斯坦奶牛乳蛋白率及体细胞数呈显著相关性(p0.05);I18-652AG位点与中国荷斯坦奶牛乳蛋白、干物质及体细胞数呈显著相关性(p0.05);I18-726AG与中国荷斯坦奶牛乳蛋白率、奶量、干物质、体细胞数均呈显著相关性(p0.05)。其次构建了牛GPAM基因过表达载体及干扰载体,转染奶牛乳腺上皮细胞后,对GPAM基因与脂代谢的相关功能进行验证。实验结果表明:在奶牛乳腺上皮细胞中,牛GPAM基因能调节脂肪代谢信号通路中关键因子AGPAT4基因的m RNA表达水平,当GPAM基因过表达时,AGPAT4基因的m RNA表达水平显著上升;当GPAM基因沉默时,AGPAT4基因的m RNA和蛋白表达水平显著下降。除此之外,牛GPAM基因过表达时,奶牛乳腺上皮细胞中的甘油三酯含量显著上升;牛GPAM基因沉默时,奶牛乳腺上皮细胞中的甘油三酯含量显著下降。实验结果表明:GPAM基因可能在调节动物脂肪代谢中起关键作用。
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S823

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