结核分枝杆菌早期分泌蛋白CFP-10的原核表达

Prokryotic expression of CFP-10 protein of Mycobacterium tuberculosis

目的构建结核分枝杆菌（MTB）早期分泌蛋白CFP-10原核表达载体并进行诱导表达。方法通过PCR技术从结核分枝杆菌H37Rv基因组中扩增出lhp基因片段，目的基因片段克隆至表达载体pET-32a（＋），构建pET-32a—CFP-10重组表达质粒，将其转化E．coliBL21（DE3），经IPTG诱导表达。结果成功地构建原核表达载体pET-32a—CFP-10，以重组体转化E．coliBL21（DE3），成功进行诱导表达。结论成功构建CFPl0的原核表达质粒，高效表达目的蛋白CFP-10，为进-步研究其免疫学功能奠定了基础。

Objective To construct recombinant plasmid containing CFP-IO gene of Mycobacteri- um tuberculosis（MTB）. Methods The gene fragment of CFP-IO was amplified by PCR from Mycobacteri- um tuberculosis H37Rv genomic DNA and cloned to pET-32a（ ＋ ） vector. The recombinant plasmid pET- 32a-CFP-lO was transformed into E. coli BL21 （DE3） and induced by IPTG. Results CFP-10 gene frag- ment was amplified from genomie DNA of Mycobacterium tuberculosis H3~ Rv strain, and the pET-32a（ ＋ ） prokaryotic recombinant plasmid was constructed successfully. The recombinant protein was expressed with the induction of IPTG. Conclusions The prokaryotic expression vector for CFP-IO was successfully constructed and the recombinant protein was highly expressed in E. coli BL21 （DE3） , which lays a foundation for its subseauent immunological function study.