【摘要】 目的　为研究结核分枝杆菌基因Rv0 90 1的功能提供材料。方法　PCR扩增Rv0 90 1基因编码序列 ,定向克隆入融合蛋白原核表达载体pGEX 1λT获得重组表达质粒 ,转化大肠杆菌后用IPTG进行诱导表达 ,通过SDS PAGE、GST 纯化试剂盒鉴定并纯化表达产物。结果　从结核杆菌H37Rv株基因组DNA中扩增出Rv0 90 1基因 ;成功构建了融合表达质粒pGEX Rv0 90 1;重组质粒经IPTG诱导后能在大肠杆菌中稳定表达 4 5kDa的GST Rv0 90 1融合蛋白。结论　本实验成功表达并纯化GST Rv0 90 1融合蛋白 ,为Rv0 90 1基因功能的研究打下了基础。更多还原

【Abstract】 Objective To construct prokaryotic expression vector as the basis for exploring the function of a unknown gene Rv0901 of Mycobacterium tuberculosis.Methods The gene encoding Rv0901 protein of Mycobacterium tuberculosis H37Rv strain was amplified by PCR and cloned into prokaryotic expression vector pGEX-1λT.The recombinant plasmid was transformed into E.coli JM109 to express the fusion protein GST-Rv0901 induced by IPTG.The expression product was analyzed by SDS-PAGE and purified by the Bulk GST Purification Module.Results The gene Rv0901 was amplified accurately from the genome DNA of H37Rv.A recombinant fused expression vector pGEX-Rv0901 was constructed.The recombinant plasmid can express the fusion protein GST-Rv0901 stably.Conclusions The 45 kDa fusion protein GST-Rv0901 was successfully expressed and purified.It provided the basis for the further study of the gene Rv0901.更多还原