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Periplasmic expression and one-step purification of urease subunit B of Helicobacter pylori

Abstract

UreB is one of the urease subunits of Helicobacter pylori and can be used as an excellent antigen candidate for H. pylori vaccination. Easy access to highly purified UreB protein, facilitate advances in therapeutic or preventive strategies. To achieve a simplified purification procedure, the present report represents a novel method of producing recombinant urease subunit B extracellularly. ureB gene from 26,695 standard strain was amplified by PCR and cloned into pET-26b(+) expression vector. UreB was expressed as a soluble, N-terminal pelB and C-terminal hexahistidine-tagged fusion protein (UreB-6His) and secreted into the periplasmic space of Escherichia coli. Expression of the recombinant UreB in E. coli BL21 (DE3) was induced by isopropylthio-β-d-galactoside (IPTG). Expression of UreB was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blot analysis using anti-His monoclonal antibody. UreB-6His protein was extracted from the periplasm by osmotic shock treatment and was purified in one step by Nickel affinity chromatography. In conclusion, the present protocol is easier to perform; more time effective and low cost than earlier methods.

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Acknowledgments

This article has been extracted from an approved research (application No. U-870244) which has financially been supported by Research Deputy of Jundishapur University of Medical Sciences, Ahvaz, Iran. The authors would like to thank Mrs. Neisi and Mrs. Lotfi from Virology Department for assistance with this work, and Mrs. Noorbehbahani and Mrs. Amuzegari from Biochemistry Department of Jundishapur University of Medical Sciences, Ahvaz, Iran for their experience in chromatography throughout this study.

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Correspondence to Amirhooshang Alvandi.

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Alvandi, A., Farajzadeh, A., Ghaforian Borojerdnia, M. et al. Periplasmic expression and one-step purification of urease subunit B of Helicobacter pylori . World J Microbiol Biotechnol 27, 969–974 (2011). https://doi.org/10.1007/s11274-010-0540-2

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Keywords

  • Helicobacter pylori
  • Urease subunit B
  • Cloning
  • Periplasmic expression