GPAM基因过表达载体的构建及其在奶牛乳腺上皮细胞中的表达

发布时间：2018-06-15 09:39

本文选题：GPAM基因 + 奶牛乳腺上皮细胞　；参考：《吉林农业大学学报》2017年02期

【摘要】：线粒体3-磷酸甘油酰基转移酶(GPAM,glycerol-3-phosphate acyltransferase)是一种具有编码蛋白质功能的基因,位于牛的26号染色体上。GPAM酶参与催化三酰基甘油和磷脂生物合成反应的第一步,继而对甘油三酯的量进行调控,是生物体内一个必不可少的酶。试验以中国荷斯坦牛RNA反转录成cDNA,继而PCR扩增出GPAM片段,并连入pBI-CMV3载体,构建了pBI-CMV3-GPAM真核表达载体,并验证了其在奶牛乳腺上皮细胞中的表达情况。结果表明:成功构建了牛GPAM真核表达载体,并在体外奶牛乳腺上皮细胞中高效表达。
[Abstract]:Mitochondrial glycerol-3-phosphate acyltransferase is a protein-encoding gene located on chromosome 26 of cattle. GPAM is the first step in catalyzing the biosynthesis of triacylglycerol and phospholipid. The amount of triglyceride is regulated, which is an essential enzyme in organism. In this experiment, the Chinese Holstein bovine RNA was inverted into cDNA. then the GPAM fragment was amplified by PCR and inserted into pBI-CMV3 vector. The eukaryotic expression vector of pBI-CMV3-GPAM was constructed, and its expression in bovine mammary epithelial cells was verified. The results showed that bovine GPAM eukaryotic expression vector was successfully constructed and highly expressed in bovine mammary epithelial cells in vitro.
【作者单位】：吉林大学动物科学学院;克莱姆森大学农业林业与生命科学学院;吉林农业大学吉林省反刍动物繁殖生物技术与健康养殖工程实验室;
【基金】：产业技术研究与开发专项(2016C032) 国家科技重大专项(2016ZX08009003-006-002) 国家“863”计划项目(2013AA102505) 国家自然科学基金项目(31372278)
【分类号】：S823

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