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

Alvandi, Amirhooshang and Farajzadeh, Ahmad. and Ghaforian Borojerdnia, Mehri. and Jelodar, Abbass. and Aryan, Ehsan. and Gholipour, Abolfazl. and Masjedizadeh, Abdolrahim. and Makvandi, Manoochehr. (2011) Periplasmic expression and one-step purification of urease subunit B of Helicobacter pylori. World Journal of Microbiology and Biotechnology, 27 (4).

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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.

Item Type: Article
Uncontrolled Keywords: Helicobacter pylori Urease subunit B Cloning Periplasmic expression
Subjects: QU Biochemistry > Cell biology and genetics
Divisions: Reserach Vice-Chancellar Department > Cellular and Molecular Research Center
Depositing User: Users 1 not found.
Date Deposited: 13 Dec 2017 06:24
Last Modified: 28 Feb 2018 06:44
URI: http://eprints.skums.ac.ir/id/eprint/6612

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