title: Molecular cloning and characterisation of the methionine sulphoxide reductase A (msrA) gene locus in Campylobacter lari organisms.
abstract: The methionine sulphoxide reductase A (msrA) gene and its adjacent genetic loci from urease-negative (UN) Campylobacter lari RM2100 and urease-positive thermophilic Campylobacter (UPTC)CF89-12 strains appear to be composed of a msrA structure gene (507 base pairs [bp]) and another five-gene cluster (approximately 6300 bp) in the same strand and direction. A primer pair (F1/R4-msrA) for polymerase chain reaction (PCR) amplification was designed to generate a product of approximately 900 bp of the msrA gene, including its adjacent genetic loci for the thermophilic Campylobacter organisms and generate an amplicon with 16 C. lari isolates (n = 4 for UN C. lari; n = 12 for UPTC). Following direct nucleotide sequencing, sequence analysis and nucleotide sequence alignment analysis, the putative full-length msrA gene from the 16 C. lari isolates showed high nucleotide sequence similarities (91.8-100%) to each other and relatively low similarity (69.3-71.8%) to three reference C. jejuni and C. coli strains. In addition, the msrA gene was transcribed in both the UPTC CF89-12 and NCTC12893 cells using reverse transcription PCR. An immunoreactively positive signal was identified in the UPTC CF89-12 and NCTC12893 cells with anti-UPTC MsrA synthetic peptide antibodies.
authors.affiliation: Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nakajima
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Matsubara
authors.affiliation: Department of Bacteriology, Northern Ireland Public Health Laboratory, Belfast City Hospital, UK.
authors.forename: J E
authors.initials: JE
authors.lastname: Moore
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Murayama
authors.affiliation: Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Matsuda
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D044863:Campylobacter lari / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers; D015723:Gene Library; D051150:Methionine Sulfoxide Reductases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D018384:Oxidative Stress; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D051150:Methionine Sulfoxide Reductases; C027219:methionine sulfoxide reductase