畜牧与兽医2018，Vol.50Issue(4)：1-6,6.
pEGFP-C1-ApoE真核表达载体的构建及在HEK-293T细胞中的表达
Construction of eukaryotic expression vector of pEGFP-C1-ApoE and its expressions in HEK-293T cells
摘要
Abstract
Apolipoprotein E is one of the important genetic factors that determine the level of cholesterol in plasma,and,as a structural protein of a variety of lipoproteins,it plays a very important role in the transport and metabolism of lipids.In order to investigate the expression of ApoE gene in eukaryocyte,the Congjiang Xiang pigs were used to clone the CDS region of ApoE gene,to construct recombination plasmid pEGFP-C1-ApoE for the gene of sus scrofa and transfect it into the HEK-293T cell.The sequence analysis showed,compared with ApoE gene of sus scrofa submission in GenBank,that eight bases mutations were identified in the ApoE gene in Congjiang Xiang pigs,seven of the above-mentioned mutations took place between C and G.Another mutation occurred at 103 of T to C,causing the serine (S) mutation to be a rare cipher subproline (P) which increased the number of rare codons from 23 to 24.The results of bioinformatics showed that the secondary structure and tertiary structures and the physicochemical properties of ApoE were changed after the mutation.The signal peptide prediction showed that a signal peptide had been found in 1-18 amino acid residues of ApoE.Both of PSOR Ⅱ Prediction and fluorescence co -localization revealed that the expression of ApoE protein was observed mainly in the cytoplasm.The present research results provide a basis for construction ApoE gene transgenic animal models.关键词
从江香猪/载体构建/HEK-293T细胞/细胞转染/基因表达Key words
Congjiang Xiang Pig/vector construction/HEK-293T Cell/cell transfection/gene expression分类
农业科技引用本文复制引用
赵佳福,嵇辛勤,杨远青,段志强,龚婷..pEGFP-C1-ApoE真核表达载体的构建及在HEK-293T细胞中的表达[J].畜牧与兽医,2018,50(4):1-6,6.基金项目
贵州省科技重大专项(黔科合重大专项字[2013]6008号) （黔科合重大专项字[2013]6008号）
省校联合基金(黔科合LH字[2014]7668) （黔科合LH字[2014]7668）
贵州大学青年基金(贵大自青基合字[2012]010号) （贵大自青基合字[2012]010号）
