融合基因Mdc-hly的构建及原核表达  

Construction and Prokaryotic Expression of Fusion Gene Mdc-hly

作　　者：卢雪梅[1] 金小宝[1] 朱家勇[1] 梅寒芳[1] 马艳[1] 王艳[1] 李小波[1] 褚夫江[1] 

机构地区：[1]广东药学院基础学院广东省生物活性药物研究重点实验室,广州510006

出　　处：《生物技术通报》2010年第4期150-155,共6页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30671832)

摘　　要：构建家蝇天蚕素-人溶菌酶(Mdc-hly)融合基因,实现Mdc-hly基因在大肠杆菌中的表达。通过RT-PCR分别扩增出家蝇天蚕素和人溶菌酶的成熟肽基因序列,再利用Gene-SOEing技术构建融合基因,将融合基因克隆至pET32a表达载体,转化E.coli BL21(DE3),经IPTG诱导得到高效表达,融合蛋白分子量约为38kD。Western blotting杂交证实了表达蛋白的抗原活性。成功构建了融合其因并进行了原核表达,为进一步的生物活性研究打下基础。The DNA fragment encoding the mature peptide of Musca domestica cecropin and human lysozyme were obtained by RT-PCR. The Mdc-hly gene was constructed in a Mdc-linker-hly format with the standard 15-amino acid linker （Gly4Ser）3 by Gene-SOEing,and the final full length product was cloned into the pET-32a vector for protein expression in Escherichia coli strain BL21（DE3）. The expression fusion protein Mdc-hly-Trx were about 38 kD and Western blotting analysis confirmed that the 38 kD protein was the fusion protein,because it was specifically recognized by mouse anti-His monoclonal antibody. The fusion gene Mdc-hly was successfully constructed and expressed,the results of which could provide foundation for further study of its biological activity.

关 键 词：家蝇天蚕素 人溶菌酶 Gene-SOEing 原核表达 

分 类 号：Q78[生物学—分子生物学]