Cloning and Gene Expression of ureG Gene as a DNA Vaccine Candidate Against Helicobacter Pylori

Helicobacter pylori is a gram-negative bacterium that has infected many human societies worldwide. the urease of H. pylori is essential for its survival in the human stomach and the ureG gene is one of the most important virulence factors.
The aim of this study is the cloning of the ureG gene in order to generate a gene vaccine against H. pylori.
In this experimental study, the DNA was extracted from helicobacter pylori. amplification of ureG gene was performed using specific primers and the PCR products were cloned into pTZ vector using T/A cloning technique. the ureG gene was then cut from the pTZ vector using the XbaI and SalI enzymes and the gene was subcloned in the pCI-neo expression vector. the pCI-neo-ureG recombinant vector was transformed into CHO cells by electroporation, and ureG gene expression was detected on a SDS-PAGE gel.
The H. pylori ureG gene was amplified and isolated by PCR successfully. the results showed that the ureG gene was cloned into pTZ and pCI-neo vectors, correctly and the pCI-neo-ureG final construct was generated. the results of insertion of final construct into CHO cells showed the 23 KDa product on SDS-PAGE gel.
The ureG gene cloned into pCI-neo expression vector has the potency of expression and production of the specific product of this gene in CHO animal cell. therefore, this gene construct can be used as a suitable candidate for the further experiments of recombinant vaccines against H. pylori in animal models.