title: Construction, expression and characterisation of recombinant molecules of the urease gene operon from a urease-positive thermophilic Campylobacter (UPTC) isolate.
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.
authors.affiliation: Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara 252-5201, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Nakanishi
authors.forename: T
authors.initials: T
authors.lastname: Nakajima
authors.forename: A
authors.initials: A
authors.lastname: Tazumi
authors.forename: K
authors.initials: K
authors.lastname: Matsubara
authors.forename: J E
authors.initials: JE
authors.lastname: Moore
authors.forename: B C
authors.initials: BC
authors.lastname: Millar
authors.forename: M
authors.initials: M
authors.lastname: Matsuda
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002167:Campylobacter / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli; D017353:Gene Deletion; D008969:Molecular Sequence Data; D009532:Nickel; D009876:Operon; D011994:Recombinant Proteins / Q000378:metabolism; D014510:Urease / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; C022838:nickel chloride; D009532:Nickel; D014510:Urease