Construction, expression and characterisation of recombinant molecules of the urease gene operon from a urease-positive thermophilic Campylobacter (UPTC) isolate.

Construction, expression and characterisation of recombinant molecules of the urease gene operon from a urease-positive thermophilic Campylobacter (UPTC) isolate. Br J Biomed Sci. 2013;70(1):15-21 Authors: Nakanishi S, Nakajima T, Tazumi A, Matsubara K, Moore JE, Millar BC, Matsuda M Abstract A recombinant molecule of the full-length urease gene operon was constructed in vitro from the Japanese urease-positive thermophilic Campylobacter (UPTC) CF89-12 isolate and expressed in Escherichia coli cells. Several large deletion recombinant variants of urease subunit genes were also constructed and expressed in E. coli cells. A positive urease reaction with the log-phase cultured E. coli JM109 cells in the NiCl2-containing medium transformed with pGEM-T vector carrying the recombinant molecule of the full-length operon was detected with isopropyl-beta-D-thiogalactoside. Among the several deletion recombinant variants, each ureA-, ureB-, ureE-, ureF-, ureG- and ureH-large deficient, only ureE-large deletion variant (63% deficient) showed a positive urease reaction (approximately 15-fold). In addition, a ureE-complete deletion recombinant variant (100% deficient) constructed also showed a positive reaction of urease (approximately 18-fold). Recombinant urease subunits A and B were immunologically identified by Western blot analysis with anti-urease alpha (A) and beta (B) raised against Helicobacter pylori. PMID: 23617093 [PubMed - indexed for MEDLINE]
Source: British Journal of Biomedical Science - Category: Laboratory Medicine Tags: Br J Biomed Sci Source Type: research

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