1
Microbiology Department, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2
Biochemistry Department, School of Basic Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Objective: Helicobacter pylori is associated with chronic gastritis, peptic ulcers, gastric adenocarcinoma and gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Antibiotic therapies do not protect from potential re-infection and have a risk for development of drug resistance. Therefore, prophylactic vaccine mediated protection against H. pylori is an attractive clinical interest. H. pylori adhesin A (HpaA) is a conserved surface lipoprotein and plays important 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 vector pET28a-hpaA was constructed, and used to transform E.coli BL21DE3. The expression of recombinant protein induced by IPTG was examined by SDS-PAGE. Western blot were used to determine immunoreactivity of rHpaA by a rabbit polyclonal antibodies against whole cell 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-PAGE demonstrated that the constructed prokaryotic expression efficiently produced rHpaA at the 1.5 mmol/L of IPTG. HpaA fusion protein was able to react with the rabbit polyclonal antibody against 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 satisfactory immunoreactivity. These results indicate that production of a specific recombinant protein is an alternative and potentially more expeditious strategy for development of H. pylori vaccine
Peerayeh, S. N., Atoofi, J., Hoseinkhani, S., & Farshchian, M. (2009). Cloning And Expression Of Helicobacter Pylori HpaA Gene. Cell Journal (Yakhteh), 11(3), 273-276.
MLA
Shahin Najar Peerayeh; Javad Atoofi; Saman Hoseinkhani; Moein Farshchian. "Cloning And Expression Of Helicobacter Pylori HpaA Gene". Cell Journal (Yakhteh), 11, 3, 2009, 273-276.
HARVARD
Peerayeh, S. N., Atoofi, J., Hoseinkhani, S., Farshchian, M. (2009). 'Cloning And Expression Of Helicobacter Pylori HpaA Gene', Cell Journal (Yakhteh), 11(3), pp. 273-276.
VANCOUVER
Peerayeh, S. N., Atoofi, J., Hoseinkhani, S., Farshchian, M. Cloning And Expression Of Helicobacter Pylori HpaA Gene. Cell Journal (Yakhteh), 2009; 11(3): 273-276.
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