Background & Objectives: Flagella and motility are important factors in bacterial colonization and pathogenicity. flaB is one of the important genes encoding the flagellar protein, which produces antibody against this gene play major role in immunogenesis. The aim o More
Background & Objectives: Flagella and motility are important factors in bacterial colonization and pathogenicity. flaB is one of the important genes encoding the flagellar protein, which produces antibody against this gene play major role in immunogenesis. The aim of the present study was to make a flaB gene construct and to evaluate its expression in eukaryotic cells as a candidate for vaccine production. Materials & Methods: flaB cloning and subcloning was done in pTZ57RT and pBudCE4.1 plasmids. Then the accuracy of the obtained clones confirmed by PCR, double enzyme digestion and sequencing. After transferring the confirmed gene construct to the animal cells, the gene expression was checked at the level of transcriptome and proteome by RT-PCR and SDS-PAGE methods. Results: PCR results showed amplification of a1563 bp segment related to flaB gene. Desired gene cloning had confirmed by PCR, double enzyme digestion and sequencing. The observation of 1563 bp and 56 kDa bands from RT-PCR and SDS-PAGE indicates the successful expression of flaB in HDF cells.Conclusion: The recombinant gene construct can express the FlaB protein in animal cells. Therefore, according to the results, this construct can be used as a candidate to produce an effective gene vaccine against Helicobacter pylori.
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