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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。
Objective: To construct eukaryotic recombinant plasmids of mycobacterium tuberculosis secreting antigen 85B. Methods: The 85B specific primers were designed and the 85B gene fragment was amplified from 85B plasmid DNA by polymerase chain reaction (PCR) and cloned into pMD18-T vector. After sequencing, it was subcloned into eukaryotic expression vector pVAX1, PCR and double digestion identified positive strains. Results: The 85B gene fragment was amplified by PCR. The correct fragment was subcloned into the eukaryotic expression plasmid pVAX1 to construct eukaryotic recombinant plasmid pVAX1-85B. Conclusion: The eukaryotic recombinant plasmid pVAX1-85B of 85B was successfully constructed.