摘要
目的在大肠杆菌中表达、纯化结核分枝杆菌Rv0173抗原,为研制新型结核病疫苗打下基础。方法将结核杆菌抗原Rv0173的全长cDNA插入到原核表达载体pGEX-4T-1中,构建成Rv0173重组质粒。将重组质粒转入大肠杆菌BL21后用IPTG进行诱导表达,通过SDS-PAGE和Western-blot鉴定重组表达蛋白。结果获得了pGEX4T-Rv0173重组子,Rv0173蛋白在BL21菌中获得表达,表达的蛋白条带大小约45KD,与预期结果相符。结论成功地对结核杆菌免疫保护性抗原Rv0173进行了基因克隆与表达,为进一步研究其在结核病新型疫苗研制中的应用奠定了基础。
Objective To express recombinant antigen Rv0173 of Mycobacterium tuberculosis(Mtb.) for development of novel TB vaccine.Methods The whole gene containing DNA sequence encoding antigen Rv0173 was synthesized and used as a template to amplify the DNA sequence by polymerase chain reaction(PCR) for expressing recombinant Rv0173 protein,containing restriction sites(BamHⅠ、XhoⅠ) on 5' and 3' end respectively.The PCR product was inserted into expression vector pGEX-4T-1 and then the recombinant plasmid was transformed into E.Coli BL21,induced by isopropylthio-β-D-galactoside(IPTG).Results The recombinant 45KD Rv0173 protein was expressed and confirmed by SDS-PAGE and Western-blot.Conclusion The recombinant TB antigen Rv0173 was expressed successfully.
出处
《中国实验诊断学》
2007年第7期914-916,共3页
Chinese Journal of Laboratory Diagnosis
基金
本课题受国家自然科学基金(30471616)
上海市重大科技攻关基金(04DZ19116)
上海市重点科研支撑条件项目(051409012)
上海市青年科技启明星计划项目(QMX01423)
上海市重点基础研究项目(05JC14052)资助
