hnRNP A2/B1基因的蛋白表达初步探讨  

Construction of an Expression Vector containing the hnRNPA2/B1 Gene

作　　者：雷祖宝[1] 操乐杰[1] 李润生[1] 李先武[2] 

机构地区：[1]安徽医科大学附属省立医院呼吸科,合肥市230001 [2]华盛顿大学生物医学工程系

出　　处：《中国肿瘤临床》2006年第15期854-857,共4页Chinese Journal of Clinical Oncology

基　　金：安徽省科技厅重点科研基金(编号:01023024;05023086);安徽省教育厅自然科学基金资助(编号:2004kj238zc)

摘　　要：目的:克隆hnRNPA2/B1基因并构建全长cDNA的原核表达载体,在大肠杆菌中进行表达。方法:以RT-PCR法从人平滑肌肌细胞总RNA中克隆hnRNPA2/B1cDNA,连接至pGEX-4T-1载体,构建重组表达质粒,转化感受态E.coliBL21进行诱导表达。结果:克隆hnRNPA2/B1基因全长cDNA序列,经DNA测序证明与GenBank一致。进而构建重组表达载体PCEX-A2/B1,在E.coli中表达出相对分子量约63kD的融合蛋白,免疫分析证实为hnRNPA2/B1蛋白。结论:成功克隆hnRNPA2/B1基因,在E.coli获得融合表达,为进一步研究其功能及应用提供基础。Objective： To prepare an expression vector containing hnRNPA2/B1 fusion protein in order to synthesize monoclonal antibody. Methods： The cDNA of hnRNPA2/B1 gene was cloned from the total RNA of human muscle cells by RT-PCR. After sequencing, the gene was cloned into the expression vector PGEX-4T-1 to construct a recombinant plasmid named PGEX-A2/B1. E. coli BL21 cells containing the expression plasmid were induced with IPTG. Results： The full-length cDNA of the hnRNPA2/B1 gene was successfully cloned and confirmed by DNA sequencing. A prokaryotic expres- sion vector for the hnRNPA2/B1 gene was constructed, and the fusion protein was expressed in E.coli after IPTG induction. Conclusion： The hnRNPA2/B1 gene was successfully cloned and a recombinant plasmid PGEX-A2/B1 containing this gene was constructed. The fusion protein expressed in E.coli was identified as hnRNPA2/B1 by Western blotting.

关 键 词：HNRNPA2/B1 基因克隆 融合蛋白 

分 类 号：R734.2[医药卫生—肿瘤]