结核分支杆菌热休克蛋白65kD在大肠杆菌中表达

目的　构建能表达结核分支杆菌热休克蛋白65kD的工程菌。方法 设计引物并PCR扩增,目的基因克隆并转化,重组子经酶切和自动测序鉴定,阳性重组子转化表达宿主菌并受化学诱导后表达重组65kD蛋白。结果 65kD蛋白编码基因约1.6kb,重组子单酶切和双酶切证明目的基因插入载体,3个测序反应覆盖99.1%目的基因;大肠杆菌表达重组65kD蛋白。结论 大肠杆菌表达系统是一种快速、简便获得单一结核分支杆菌抗原的途径。

Expression of 65 kD Mycobacterial heat shock protein in E. coli

Objective To construct an engineering strain that can express 65kD mycobacterial heat shock protein. Methods The gene encoding the Mycobacterium tuberculosis 65kD antigen was amplified using PCR and then was cloned in the plasmid. The recombinant plasmid was transformed into E. coli , and then was screened using enzyme digestion and sequencing. The screened recombinant plasmid was transported into E. coli , subsequently the recombinant E. coli was induced by IPTG to express 65kD protein. Results The gene encoding 65kD protein is 1.6kb. Base pairs of DNA sequencing were about 99.1% total gene. The constructed recombinant E. coli can produce 65kD protein of M. tuberculosis . Conclusion E. coli expression system is a rapid and simple means to obtain recombinant antigen of M. tuberculosis .