湖北医药学院学报2016，Vol.35Issue(4)：350-354,5.DOI:10.13819/j.issn.1006-9674.2016.04.005
结核杆菌H37Rv菌株蛋白Rv0117和Rv3379c的表达纯化及功能验证
The Expression, Purification and Functional Verification of Mycobacterium Tuberculosis H37Rv Strain Protein Rv0117 and Rv3379c
摘要
Abstract
Objective To express and purify the Mycobacterium tuberculosis H37Rv strain Rv0117 and Rv3379c protein,and to verify the activation function to γδT cells in peripheral blood mononuclear cells of tuberculosis patients.Methods We extracted the genomic DNA of H37Rv strain,the Rv0117 and Rv3379c gene were amplified by PCR.By restrict digestion of Rv0117 and Rv3379c gene,they were connected to the pET30 vector.The recombinant was transformedinto expression host E.coli BL21.The expression was induced by IPTG and was identified by polyacrylamide gel electrophoresis and Western blot.The activation function to γδT in the peripheral blood mononuclear cells of tuberculosis patients was detected by flow cytometry.Restults We successfully constructed prokaryotic expression vector pET30a-Rv0117 and PET30a-Rv3379c,and the expression and purification of Rv0117 and Rv3379c protein were further confirmed by Western blot.The Rv0117 and Rv3379c protein could activate γδT cells in the peripheral blood mononuclear cells of tuberculosis patients.Conclusion The expression and purification of Rv0117 and Rv3379c protein would establish the foundation for the development of new tuberculosis vaccine-based on γδT cells.关键词
结核杆菌/蛋白表达/γδT细胞Key words
Tuberculosis/Protein expression/γδT cells引用本文复制引用
王晓雯,晋兰,赵俊杰,郗雪艳..结核杆菌H37Rv菌株蛋白Rv0117和Rv3379c的表达纯化及功能验证[J].湖北医药学院学报,2016,35(4):350-354,5.基金项目
国家自然科学基金面上项目(31370890) （31370890）
