结核分支杆菌Mtb8.4抗原原核表达质粒的构建与鉴定(英文)  

Construction of prokaryotic recombinant plasmid encoding Mtb8.4 gene of M.tuberculosis and identification

作　　者：李素华[1] 徐建国[2] 钟森[3] 李晖[3] 

机构地区：[1]温州医学院附属育英儿童医院,浙江温州325000 [2]温州市第三人民医院ICU,浙江温州325027 [3]泸州医学院附属医院感染科,四川泸州646000

出　　处：《中国现代医学杂志》2005年第7期966-968,共3页China Journal of Modern Medicine

摘　　要：目的构建结核杆菌低分子质量抗原M tb8.4基因的原核表达重组质粒,为其蛋白质的表达及免疫学研究打下基础。方法PC R扩增目的基因,克隆到质粒pET-H is T7启动子的下游,转化大肠杆菌T O P10F',进行重组子的筛选、鉴定。结果M tb8.4基因正向插入表达载体中,测序证实具有正确的读码框,基因序列与文献报道一致。结论重组原核表达质粒pET-H is-M tb8.4构建成功。To construct prokaryotic expression recombinant plasmid encoding Mtb8.4 gene of M.tuberculosis. The gene encoding protein Mtb8.4 was amplified by PCR technique. Clone the gene into the downstream of the T7 promoter pET-His, then transformed into E.coli TOP10F′. Positive clone was screened using double digestion and polymerase chain reaction, and farther identified by nucleotide sequence. Mtb8.4 gene was successfully inserted into the prokaryotic plasmid pET-His. The reading frame and gene sequence of recombinant plasmid were correct by sequencing reaction. [Conclusions] Recombinant plasmid pET-His-Mtb8.4 is successfully constructed, It will provide further study for the expression Mtb8.4 protein and its immunological effect, and a potential candidate for inclusion in a subunit vaccine against tuberculosis(TB).

关 键 词：结核分支杆菌 低分子质量抗原 克隆 原核表达质粒 

分 类 号：R521.01[医药卫生—内科学]