人肝再生增强因子融合蛋白表达载体的构建

目的 :克隆人肝再生增强因子的cDNA并构建谷胱甘肽巯基转移酶 -人肝再生增强因子 (GST -hALR)融合蛋白表达载体。方法 :根据人肝再生增强因子核苷酸序列 ,从人胎肝cDNA文库中扩增到hALRcDNA ,亚克隆于pUC18载体 ,核苷酸序列测定证实为hALR ;将hALRcDNA亚克隆于 pGEX - 4T质粒 ,转化大肠杆菌JM10 9,筛选阳性克隆酶切鉴定。结果 :重组阳性克隆经酶切鉴定证实重组片段已正确插入相应酶切位点 ,片段与PCR扩增片段大小相同 ,碱基序列正确。结论 :GST -hALR融合蛋白表达载体的成功构建 ,为获得较大量基因工程hALR产品用于重症肝炎治疗打下基础

CONSTRUCTION OF EXPRESSION VECTOR OF GST-hALR FUSION PROTEIN

Objective:To clone the cDNA of human augmenter of liver regeneration and construct the expression vector of GST-hALR fusion protein.Methods:According to the nuclear acid sequence of human augmenter of liver regeneration,we cloned the genes with PCR from human fetal liver cDNA library,then the genes were subcloned into the pUC18 vector,the sequence of hALR cDNA was proved to be correct by sequencing.Then the hALR gene was insected into the MCS of the fusion gene vector of pGEX-4T,the recombinant of vector was transformed into E.coli.JM 109,the positive clones were selected and plasmid DNA was digested with restriction endonuclease.Results:The hALR cDNA was successfully inserted into pGEX-4T,and the sequence of hALRcDNA is correct.Conclusions:Successfully constructing expression vector of GST-hALR fusion protein,can provide an essential preparation for obtaining a large quantity of recombinant human augmenter of liver regeneration to study the treatment of severe hepatitis.