Cloning and study of expression of Helicobacter Pylori flaB gene in eukaryotic system for vaccine design

Subject Areas : Molecular Microbiology

Pouya  Khodadadi ¹ (Department of microbiology, Azad university Jahrom branch. Jahrom, Iran.)
Mohammad  Kargar ² (Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran)
Mahdi  Bijanzadeh ³ (Department of Medical Genetic, Ahvaz Jundishapur University of Medical Sciences, Ahvaz,)
Abbas  Doosti ⁴ (Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran)
Shapoor  Aghaei ⁵ (Department of Internal Medicine, Yasuj University of Medical Sciences, Yasuj, Iran)

Received: 2022-12-26 Accepted : 2023-05-15 Published : 2023-06-05

Keywords: RT-PCR, cloning, recombinant vaccine, Keywords: Helicobacter pylori,

Abstract :

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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