结核分枝杆菌蛋白Hsp16.3的基因克隆以及表达

目的克隆结核分枝杆菌蛋白Hsp16.3的基因，在大肠杆菌中进行表达。方法以结核分枝杆菌H37Rv基因组DNA为模板，通过PCR方法对Hsp16.3的基因进行扩增，以pET30a为载体构建重组质粒，再转化到表达宿主菌BL21(DE3)中，以IPTG诱导表达。聚丙烯酰胺凝胶电泳(SDS PAGE)分析蛋白表达形式。结果构建了具有正确基因序列的Hsp16.3重组表达质粒，重组蛋白在大肠杆菌BL21(DE3)中表达。结论目的基因克隆入宿主菌中并表达成功，为深入研究该蛋白的生物学、免疫学活性奠定了基础。

Cloning and expression of the Mycobacterium tuberculosis gene protein Hsp16. 3

The aim was to clone and express the gene coding for Mycobacterium tuberculosis antigen Hsp16.3,and to establish a basis for the further research of biology and immunology.The gene coding for Hsp16.3 was amplified by polymerase chain reaction（PCR）,and then the gene was inserted to vector pET30a to construct the recombinant plasmid.The recombinant plasmid was transformed into expressive vector E.coli BL21（DE3） and was induced with IPTG.The presence of recombinant protein in the expression vector was analyzed by SDS-PAGE.The results indicated that the recombinant plasmid with the correct target gene was constructed,and the recombinant protein was expressed in E.coli BL21（DE3）.In conclusion the target gene was cloned into the host bacterium and expressed correctly,and these results would establish the basis for the further research in biology and immunology of Hsp16.3.