结核分支杆菌抗原85B真核表达质粒的构建

目的:构建结核分支杆菌分泌性抗原85B的真核重组表达质粒。方法:设计85B的特异引物,采用多聚酶链反应(PCR)技术从含85B基因质粒DNA中扩增85B基因片段,克隆至pMD18-T载体,经测序分析后,亚克隆至真核表达载体pVAX1,PCR和双酶切鉴定阳性菌株。结果:PCR扩增获得85B的基因片段,测序获得的正确片段被亚克隆至真核表达质粒pVAX1构建真核重组表达质粒pVAX1-85B。结论:成功构建了85B的真核重组表达质粒pVAX1-85B。

Construction of Recombinant Eukaryotic Expression Plasmid Containing Mycobacterium Tuberculosi Ag85B

Objective To construct a eukaryotic expression plasmid containing gene of Mycobacterium tuberculosi antigen 85B.Methods The DNA fragment encoding Ag85B was amplified by PCR with designed primer from the plasmid which contains the gene of 85B and cloned into pMD18-T vector,The right fragment of 85B gene determined by sequencing was subcloned into the eukaryotic expression plasmid pVAX1 and the recombinant expression plasmid pVAX1-85B was characterized by PCR and digestion with XbaⅠand BamHⅠ.Results The DNA fragment encoding Ag85B was amplified by PCR and the 85B gene fragment with right sequence was subcloned into pVAX1 to construct the recombinant eukaryotic expression plasmid pVAX-85B.Conclusion the recombinant eukaryotic expression plasmid pVAX-85B was successfully constructed.