结核分枝杆菌保护性抗原 TB10�4基因的克隆和真核表达载体的构建∗

摘要

结核分枝杆菌保护性抗原TB10�4属于结核分枝杆菌早期分泌蛋白中的ESAT⁃6家族，其免疫原性等与ESAT⁃6相当或者更佳，具有十分重要的研究价值。利用分子生物学技术克隆该基因，并将其与真核表达载体pEGFP⁃N1连接，构建重组质粒pEGFP⁃N1⁃TB10�4，为TB10�4基因疫苗的研究奠定基础。结果表明：成功克隆到TB10�4基因，测序结果与GenBank（ NP＿214802�1）中已发表的TB10�4基因序列同源性为100％。%Mycobacterium tuberculosis protective antigen TB10�4 belongs to ESAT⁃6 family of Myco⁃bacterium tuberculosis early⁃secreted protein, and it works equally or better in immunogenicity than ESAT⁃6. In this study, the gene was cloned by molecular biological technique. Compared with the published TB10�4 gene sequence in GenBank ( NP_214802�1) , their homology was found as high as 100. Then it was connected with eukaryotic expression vector pEGFP⁃N1 to build recombinant plasmid pEGFP⁃N1⁃TB10�4. The study provides experimental data for application of TB10�4 gene in Mycobacterium tuberculosis diagnosis and vaccine antigens.