Cloning and Expression of Recombinant Helicobacter pylori Urease A and B Subunits as a Putative Vaccine

Authors

  • Mohsen Karimi
Abstract:

Helicobacter pylori infection is among the most prevalent infections in the world involving more than half the adult population. H. pylori infection results in active chronic gastritis, peptic ulcers and enhances the risk of gastric malignancies. It is of utmost importance to prevent H. pylori infection particularly in highly prevalent countries including Iran. The urease holoenzyme produced by the entire Helicobacter species is essential for their virulence such that urease-negative mutant strains are unable to colonize the gastric lumen of various animal models. Furthermore, urease has shown to be an effective immunogen. Despite the fact that urease is considered among the very conserved genes of this pathogen, our molecular studies have shown that H. pylori strains obtained from Iranian patients vary considerably from those of other populations particularly the Western strains. Therefore, in order to develop a putative vaccine against H. pylori infection for the Iranian population, we have PCR-amplified and cloned the A and B subunits of this gene from a local H. pylori strain. Following identity confirmation, it was subcloned into a pET expression vector under the control of T7 promoter. The resulting plasmid was transformed into E. coli BL21-DE3 strain. Laboratory scale culture of the resulting transformants was analyzed by SDS-PAGE and Western blotting techniques. This analysis confirmed the expression of the A and B subunit of H. pylori urease protein up to 25% of the total cellular protein.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

cloning and expression of recombinant helicobacter pylori urease a and b subunits as a putative vaccine

helicobacter pylori infection is among the most prevalent infections in the world involving more than half the adult population. h. pylori infection results in active chronic gastritis, peptic ulcers and enhances the risk of gastric malignancies. it is of utmost importance to prevent h. pylori infection particularly in highly prevalent countries including iran. the urease holoenzyme produced by...

full text

Cloning, Expression and Characterization of Recombinant Exotoxin A-Flagellin Fusion Protein as a New Vaccine Candidate against Pseudomonas aeruginosa Infections

Background: Infections due to Pseudomonas aeruginosa are among the leading causes of morbidity and mortality in patients who suffer from impaired immune responses and chronic diseases such as cystic fibrosis. At present, aggressive antibiotic therapy is the only choice for management of P. aeruginosa infections, but emergence of highly resistant strains necessitated the development of novel alt...

full text

جداسازی و همسانه‌سازی ژن ureE هلیکوباکتر پیلوری در ناقل بیانی pIRES2-DSRed به‌منظور ایجاد واکسن ژنی

Background and Aim: As one of the factors of gastric ulcers and cancer, Helicobacter pylori can live in the acidic environment of stomach for many years due to having urease enzyme. This enzyme requires Ni2+ and a group of auxiliary proteins such as ureE for its catalytic activity. Urease is not only a requisite factor to colonize the Helicobacter pylori but it is also pathogenic with diff...

full text

Cloning, Expression, and Refolding of PPE17 Protein of Mycobacterium Tuberculosis as a Promising Vaccine Candidate

Background: Tuberculosis as a global health problem requires special attention in the contexts of prevention and control. Subunit vaccines are promising strategies to protect burdens of tuberculosis via increasing the BCG protection. The present study aimed to design a vaccine study by means of high-throughput expression and correct refolding of recombinant protein PPE17. Methods: We aimed to c...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 5  issue 4

pages  107- 111

publication date 2001-04

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023