Aim: To clone ureB gene of H.pylori and construct its gene vaccine.
Methods: ureB gene was amplified by PCR from genome of H. pylori 11637 strain and subcloned into pMD18-T vector. The vector digested with restriction enzyme (Sal I and Bgl II) was inserted into pTCAE and transformed into E. coli DH5alpha. The positive recombinant plasmid identified by digesting with restriction enzyme (Sal I and Xho I) and sequencing named pT-ureB. The pT-ureB was transfected into CHO cells by electroporation method. The expression of UreB protein was detected by Western blot.
Results: The pT-ureB was obtained by cloning and recombinant DNA technique. The Western blot analysis showed that the expression of UreB protein (M(r)approximately 62,000) was detected in culture supernatant of CHO cells following transfection with pT-ureB.
Conclusion: UreB DNA vaccine of H. pylori was successfully constructed. The expression of UreB protein can be detected in culture supernatants of transfected CHO cells.