title: [Construction and Expression of Vector with mip/flaA Advantages Epitope Genes of Legionella pneumophila].
abstract: OBJECTIVE
  To generate and express fusion vector with mip/flaA advantages epitope genes of Legionella pneumophila by select mip and flaA advantages epitope genes for future research on Legionella pneumophila protein 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 and T4 ligase 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, the mip/flaA two advantages associated epitope fusion proteins were also successfully expressed.
  
  
  CONCLUSIONS
  DNA Star software and Expasy online analysis system can successfully predict antigenic epitopes for Legionella pneumophila Mip and FlaA. And prokaryotic expression vector pET-mip/flaA with advantages epitope genes has been successfully constructed and efficiently expressed.
authors.forename: Jin-lei
authors.initials: JL
authors.lastname: He
authors.forename: Ran
authors.initials: R
authors.lastname: Peng
authors.forename: Jun-rung
authors.initials: JR
authors.lastname: Zhang
authors.lastname: Yu Ze-ying
authors.forename: Na-li-sha
authors.initials: NL
authors.lastname: Li
authors.forename: Yan-ju
authors.initials: YJ
authors.lastname: Gong
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.forename: Da-li
authors.initials: DL
authors.lastname: Chen
authors.forename: Jian-ping
authors.initials: JP
authors.lastname: Chen
mesh_terms: D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D000939:Epitopes / Q000235:genetics*; D004926:Escherichia coli; D005408:Flagellin / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors; D016952:Legionella pneumophila / Q000235:genetics*; D019696:Peptidylprolyl Isomerase / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D005408:Flagellin; C067138:flaA protein, bacteria; C512922:Mip protein, Legionella pneumophila; D019696:Peptidylprolyl Isomerase