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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。
Objective To construct the recombinant plasmid of M tb8.4 gene of Mycobacterium tuberculosis low molecular weight antigen and lay the foundation for its protein expression and immunological study. Methods The target gene was amplified by PCR and cloned into the downstream of plasmid pET-H is T7 promoter. The recombinant plasmid was transformed into E. coli T O P10F ’for screening and identification. Results The M tb8.4 gene was inserted into the expression vector in forward direction, and the sequence was verified to be correct by sequencing. Conclusion The recombinant prokaryotic expression plasmid pET-H is-M tb8.4 was constructed successfully.