Cloning and Expression of Helicobacter pylori HpaA Gene

Helicobacter pylori is associated with chronic gastritis, peptic ulcers, gastric adenocarcinomaand gastric mucosa-associated lymphoid tissue (malt) lymphoma. antibiotictherapies do not protect from potential re-infection and have a risk for development of drugresistance. therefore, prophylactic vaccine mediated protection against h. pylori is an attractiveclinical interest. h. pylori adhesin a (hpaa) is a conserved surface lipoprotein and playsimportant roles in the pathogenesis of infection. in this study the recombinant protein (rhpaa)was over-expressed in e.coli.materials and methods: the hpaa gene was amplified by pcr. prokaryote expression vectorpet28a-hpaa was constructed, and used to transform e.coli bl21de3. the expressionof recombinant protein induced by iptg was examined by sds-page. western blot wereused to determine immunoreactivity of rhpaa by a rabbit polyclonal antibodies against wholecell of h. pylori.results: the hpaa gene nucleotide sequence in the recombinant plasmid vector of pet-28-a-hpaa was consistent with that of h.pylori hpaa as published in the genbank. sds-pagedemonstrated that the constructed prokaryotic expression efficiently produced rhpaa at the1.5 mmol/l of iptg. hpaa fusion protein was able to react with the rabbit polyclonal antibodyagainst whole cells of h. pylori.conclusion: a prokaryotic expression system pet-28a-hpaa-bl21 with high efficiency of h.pylori hpaa gene was successfully established and the hpaa fusion protein showed satisfactoryimmunoreactivity. these results indicate that production of a specific recombinant proteinis an alternative and potentially more expeditious strategy for development of h. pylori vaccine.