Construction and Expression of Vector withmip/flaAAdvantages Epitope Genes ofLegionella pneumophila

Objective To generate and express fusion vector withmip/flaAadvantages epitope genes of Legionella pneumophila by select mip and flaAadvantages epitope genes for future research onLegionella pneumophilaprotein vaccine. Methods Following analysis of secondary structure and surface properties such as: physical and chemical properties, hydropathy, plasticity, antigen index and extracellular domain of Mip and FlaA proteins by bioinformatics methods, the region which active epitope may exist was selected as advantages epitope region. Then, the recombinant plasmid pET-mip, pET-flaA and pET-mip/flaA with advantages epitope genes were constructed by PCR amplification andT4ligase connection, and induced the expression in E.coli. Results Many potential antigenic epitopes in Mip and FlaA were identified, and the selected advantages epitope regions were cloned and expressed successfully. Moreover, themip/flaAtwo advantages associated epitope fusion proteins were also successfully expressed. Conclusion DNA Star software and Expasy online analysis system can successfully predict antigenic epitopes forLegionella pneumophila Mip and FlaA. And prokaryotic expression vector pET-mip/flaAwith advantages epitope genes has been successfully constructed and efficiently expressed.