结核分枝杆菌19-kDa蛋白在大肠杆菌中的表达  

Expression of 19-kDa Protein of Mycobacterium Tuberculosis in Escherchia Coli

作　　者：韩慧新[1] 李邦印[1] 吴雪琼[1] 张灵霞[1] 

机构地区：[1]中国人民解放军总医院第二附属医院结核病研究室,北京市100091

出　　处：《中国全科医学》2006年第13期1057-1059,共3页Chinese General Practice

基　　金：国家自然科学基金资助项目(30471648)

摘　　要：目的 通过基因工程技术获得重组结核分枝杆菌19-kDa蛋白。方法 应用PCR技术扩增结核分枝杆菌H37 Rv19-kDa DNA序列；以质粒pET24b为表达载体，构建19-kDa重组质粒，然后转化大肠杆菌BL21（DE3）；在异丙基硫代-13-D-半乳糖苷（IPTG）诱导下，分别对不同诱导时间的表达产物进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE），凝胶经考马斯亮蓝染色检测蛋白。结果重组质粒pET24b-19-kDa测序表明与报道的序列相同。它在大肠杆菌BL21（DE3）细胞内以可溶性形式表达。不同IPTG诱导时间实验表明重组结核分枝杆菌19-kDa蛋白诱导3～4h重组蛋白在大肠杆菌中的表达量最高。结论 pET24b-19-kDa大肠杆菌工程株可高表达结核分枝杆菌重组19-kDa蛋白。Objective To obtain recombinant 19 - kDa protein from Mycobacterium tuberculosis by using gene engi- neering technology. Methods The gene coding 19 - kDa protein inserted into a expression vector pET24b, and then transferred E. coli BL21 （ DE3 ）. The expressed product was identified by SDS - PAGE, Plasmid pET24b containingl9 - kDa gene was transferred into competent Escherichia coli BL21 （ DE3 ） and 19 - kDa gene was over expressed by the inducement of IPTG. Resuits The recombinant 19 - kDa protein was expressed in soluble form in E. coli. When the culture was induced for 3 - 4 hours, the recombinant 19 - KDa protein was produced in highest quantity . Conclusion Escherichia coli containing recombinant plasmid pET24b - 19 - kDa gene could express recombinant 19 - KDa protein in high level.

关 键 词：结核 分枝杆菌 19-kDa 克隆 基因表达 

分 类 号：R378.911[医药卫生—病原生物学]