ZHX2-GST融合蛋白原核表达载体的构建和鉴定

摘要

Objective: To construct the prokaryotic expression vector of GST-ZHX2 fusion gene in Esche-richia coli, and assay the GST-ZHX2 fusion protein. Methods:ZHX2 gene RNA was extracted from normal human liver tissue, reverse transcriptased and amplified into full length cDNA by RT-PCR, cloned into the GST tagged prokaryotic expression vector PGEX-4-1. The fusion gene GST-ZHX2 -was expressed in E-coli. BL 2KDE3) induced by IPTG and the GST-ZHX fusion protein was assayed by SDS-PAGE. Results: The sequence of the cloned ZHX2cDNA was corrected and the molecular weight of expression production assayed by SDS-PAGE -was 118 ku, coincided -with GST-ZHX2. Conclusion: The prokaryotic expression vector of GST-ZHX2 fusion gene is constructed successfully, and the fusion protein can be used in clinical and experiment research.%目的:构建编码ZHX2-GST融合基因的原核表达载体,在大肠杆菌中表达并对融合蛋白(ZHX2-GST)进行鉴定.方法:RT-PCR提取正常肝组织的RNA,然后扩增出ZHX2基因cDNA全长,克隆至含有GST标签的原核表达载体PGEX-4-1中,转化到Ecoli.BL21(DE3),IPTG诱导大肠杆菌表达ZHX2-GST蛋白,电泳分离鉴定.结果:测序结果显示克隆的ZHX2 cDNA序列正确,SDS-PAGE鉴定表达产物分子量118 ku,与ZHX2-GST分子量相符.结论:成功构建了编码ZHX2基因的ZHX2-GST原核表达载体,融合蛋白可用于临床及实验研究.