TRANSFERRING OF THE UreB GENE OF Helicobacter pylori INTO CUCUMBER (Cucumis sativus L.) PLANT

Helicobacter pylori is a gram-negative bacterium with broad distribution and has been identified as the most important pathogenic agent of peptic ulcer and gastric cancer. Urease beta subunit (UreB) has been identified as the most effective antigen against H. pylori. Because of the several advantages of plant expression systems for reproduction of edible vaccines, in this study, the urease beta subunit (UreB) gene that was fused to several sequences such as CPP, Extensin, Kozak, His tag and protease thrombin site was transferred into Cucumber (Cucumis sativus L.) genome via Agrobacterium tumafaciens mediated transformation. In vitro regeneration from cotyledon explants of cucumber was optimized in MS medium supplemented by various concentrations of NAA and BAP hormones in a factorial experiment based on completely randomized design at three replications. The putative transgenic cucumber plants were regenerated on the MS medium containing 0.3 mgL-1NAA (1-Naphthalene acetic acid), 3 mgL-1BAP (6-Benzyl amino purine) hormones and 50 mg/l Kanamycin antibiotic. The presence and expression of the UreB gene in transgenic plants was confirmed by PCR analysis using specific UreB primers and RT- PCR analysis, respectively. SDS-PAGE analysis showed a band about 67 KDa according to UreB in transgenic plants.