结核分枝杆菌pcHSP65真核表达载体的构建及序列测定

Construction of Eukaryotic Expression Vector pcHSP65 of Mycobacterium Tuberculosis and Its DNA Sequencing

目的 :构建以结核分枝杆菌热休克蛋白 6 5kU基因为基础的核酸疫苗。方法 :采用聚合酶链反应从结核杆菌H37Rv株基因中 ,扩增出HSP6 5的编码基因 ,经限制性核酸内切酶消化后 ,插入真核表达载体pcDNA3.1( )的相应酶切位点 ,并将此重组质粒进行序列测定。结果 :重组质粒的插入基因经序列测定证实为结核分枝杆菌HSP6 5。结论 :以HSP6 5编码基因为基础的真核表达载体构建成功 。

Objective: To construct nucleotide vaccine based on HSP65 gene of mycobacterium tuberculosis. Methods: The gene encoding heat shock protein 65 kilodalton was amplified from genome of mycobacterium tuberculosis H37Rv strain by polymerase chain reaction (PCR) and correctly inserted into corresponding sites of eukaryotic expression vector pcDNA3.1( ) after restriction endonuclease digestion. Results: The gene fragment inserted into the vector pcDNA3.1( ) was the HSP65 gene of mycobacterium tuberculosis H37Rv strain, which was confirmed by partial nucleotide sequencing. Conclusion :The successful construction of recombinant eukaryotic expression vector based on HSP65 gene of mycobacterium tuberculosis,lays the foundation for further researching in prevention and treatment of tuberculosis.