Genetic heterogeneity of urease gene loci in urease-positive thermophilic Campylobacter (UPTC)

Usui, Kaori; Iida, Haruna; Ueno, Hikaru; Sekizuka, Tsuyoshi; Matsuda, Motoo; Murayama, Ohoshi; Millar, B. Cherie; Moore, John E.

doi:10.1016/j.ijheh.2005.09.010

Cited by 4 publications

(9 citation statements)

References 26 publications

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“…In addition, the second ureasepositive organisms of C. sputorum biovar paraureolyticus among the genus Campylobacter were identified from bovine feces and human diarrhea (On et al 1998). Consequently, our research group has already clarified the molecular characteristics of urease gene operons from both of the two Campylobacter taxa (UPTC CF89-12 and C. sputorum biovar paraureolyticus LMG 17591) in previous (Usui et al 2006;Kakinuma et al 2007;Kakinuma et al 2008) and present studies.…”

Section: Discussionmentioning

confidence: 81%

“…The libraries were screened by colony hybridization with a probe specific for ureB, prepared using a degenerate ureB primer pair of u3f (5′-CAT ACA GAT ACT TTA AAY GA-3′) and u3r (5′-ACV GGY TSH GGD GTN GGR AT-3′). This primer pair has been designed in silico by inclusion of bacterial and jack bean urease structural genes and their deduced amino acid sequences according to Usui et al (2006).…”

Section: Methodsmentioning

confidence: 99%

“…Regarding the urease genes of UPTC, our group has already reported the genetic heterogeneity of urease gene loci in UPTC isolates obtained from different sources and in various parts of Europe and in Japan by using degenerate PCR primers designed in silico, based on sequence information of the two urease structural genes (ureA and ureB) for UPTC (Usui et al 2006). In addition, we have also described the cloning, sequencing, and characterization of a urease gene operon from the Japanese UPTC isolate, CF89-12, using a genomic DNA library construction procedure and phylogenetic analysis, based on nucleotide sequence information of both the urease structural and accessory genes (Kakinuma et al 2007).…”

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confidence: 99%

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Molecular analysis and characterization of a urease gene operon from Campylobacter sputorum biovar paraureolyticus

et al. 2011

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When recombinant plasmid DNA from a genomic DNA library and inverse PCR products of Campylobacter sputorum biovar paraureolyticus LMG17591 strain were analyzed, an approximate 6.5-kb pair region, encoding a urease gene operon, was identified. Within the operon, seven closely spaced and putative open reading frames for ureG, ureH(D), ureA, ureB, ureC, ureE, and ureF were detected in order. A possible overlap was detected between ureG and ureH(D), ureH(D) and ureA, and ureE and ureF. In addition, two putative promoter structures, probable ribosome-binding sites and a putative ρ-independent transcriptional terminator structure were identified. The urease gene operon transcription in the cells was confirmed by the reverse transcription-PCR analysis. A neighbor-joining tree constructed based on the nucleotide sequence information of urease genes showed that C. sputorum biovar paraureolyticus formed a cluster with Arcobacter butzleri, urease-positive thermophilic Campylobacter and some Helicobacter spp., separating those from the other urease-producing bacteria, suggesting a commonly shared ancestry among these organisms.

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Section: Discussionmentioning

confidence: 81%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Molecular analysis and characterization of a urease gene operon from Campylobacter sputorum biovar paraureolyticus

et al. 2011

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“…Culture and genomic DNA preparation were carried out as described previously. 15 Amplification, TA cloning and sequencing of the amplicons were carried out using procedures described previously. 15 Primer pairs were constructed for the amplification of the promoter region, the structural genes, the accessory genes of UPTC urease and a partial and putative open reading frame (ORF), including a putative transcriptional terminator (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…16 Nucleotide sequencing of the amplicons was performed as described previously. 15 Sequence analysis was carried out using the Genetyx-Mac (version 9; Genetyx Co., Tokyo, Japan) computer software. For accuracy, multiple TA-cloned PCR products were sequenced repeatedly.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterisation of urease genes from urease-positive thermophilic campylobacters (UPTC)

Kakinuma

Sekizuka

et al. 2008

British Journal of Biomedical Science

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This study aims to clarify the molecular characteristics of the urease gene operon from urease-positive thermophilic campylobacters (UPTC) obtained from different sources and in various countries. Sequence heterogeneity was observed for the promoter structures at the -35-like region among the 12 isolates examined. The most probable TTG start codon was suggested for the ureB and ureH genes, and for the ureA, E, F and G genes, ATG was suggested among all the isolates examined. Overlap was detected between ureA and ureB and between ureB and ureE among all the isolates examined. UPTC is the first example of an overlap between the two structural genes ureA and ureB. When the completely sequenced open reading frames (ORFs) for ureE, ureF, ureG and ureH were identified, non-coding regions between ureE and ureF, ureF and ureG, and ureG and ureH were also demonstrated. All six start codons of the six urease genes were demonstrated to be preceded by Shine-Dalgarno sequences among all the isolates examined. The Cys-His sequence corresponding to urease active sites were aligned perfectly and fully conserved among the three UPTC isolates examined. A putative and intrinsic p-independent transcriptional terminator was identified to be identical among all the isolates examined. A partial and putative ORF of about 200 bp in length showing high sequence similarity to GTP cyclohydrolase I was observed downstream of ureH.

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Biochemical characterisation of urease from urease-positive thermophilic Campylobacter (UPTC)

Tazumi

Nakajima

Sekizuka

et al. 2012

British Journal of Biomedical Science

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This study aims to characterise biochemically urease from an atypical Campylobacter lari, namely urease-positive thermophilic Campylobacter (UPTC). Urease was purified from cells of a Japanese UPTC isolate (CF89-12) using phenyl-Sepharose chromatography. Two protein components (estimates molecular masses 24 kDa and 61 kDa) were obtained that appeared to be structural proteins of urease (subunits A and B), and these were fractionated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (PAGE). The native molecular weight for the final purified UPTC urease was estimated to be approximately 186,000 Da which is close to the calculated molecular weight (182,738 Da) based on all six open reading frames of UPTC CF89-12 urease genes (ureA, B, E, F, G and H), as described previously. Moreover, an active band was observed on phenol red staining after a nondenaturing native PAGE of the crude extract from the UPTC cells. In addition, the purified urease of UPTC CF8912 showed enzyme activity over a broad pH range (pH 6-10), with maximal activity at pH 8.0. The urease was also stable against heat treatment, with almost no loss of enzyme activity seen following 60-min incubation at temperatures of 20-60 degrees C. Urease subunits A and B were identified immunologically by Western blot analysis with rabbit anti-urease alpha (A) and beta (B) raised against Helicobacter pylori.

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Genetic heterogeneity of urease gene loci in urease-positive thermophilic Campylobacter (UPTC)

Cited by 4 publications

References 26 publications

Molecular analysis and characterization of a urease gene operon from Campylobacter sputorum biovar paraureolyticus

Molecular analysis and characterization of a urease gene operon from Campylobacter sputorum biovar paraureolyticus

Molecular characterisation of urease genes from urease-positive thermophilic campylobacters (UPTC)

Biochemical characterisation of urease from urease-positive thermophilic Campylobacter (UPTC)

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