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

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。

Construction of GPAM Gene Overexpression Vector and Its Expression in Mammary Epithelial Cells of Dairy Cows

Mitochondrial 3-glycerol phosphate acyltransferase (GPAM and glycerol-3-phosphate acyltransferase) is a gene encoding protein function,located in 26 chromosome in cattle.GPAM enzyme catalyzes three acylglycerol and phospholipids biosynthetic reactions as a first step,and adjusts and controls the amount of triglycerides,which is an essential enzyme in the body.In this experiment,RNA of Chinese holstein was reversely transcribed into cDNA,GPAM fragments were obtained through PCR amplification,then the fragments were connected to pBI-CMV3 cartier,and finally pBI-CMV3-GPAM eukaryotic gene expression vector was constructed.This experiment verifies the expression of cow mammary gland epithelial cells in vitro.The results show that we successfully built GPAM expression vector of the cow,and the vector efficiently expressed cow mammary gland epithelial cells in vitro.