Full-length tlyA gene and its adjacent genetic loci from the urease-positive thermophilic Campylobacter (UPTC) CF89-12 [approximately 15,000 base pairs (bp) in length], as well as a reference strain Campylobacter lari RM2100 (approximately 9,000 bp), were analyzed. The possible open-reading frame of tlyA from UPTC CF89-12 was shown to have 720 bp with a calculated molecular mass of approximately 26.7 kDa. Using a primer pair designed in silico, a total of approximately 1.1 kbp consisting of putative promoter region, structural gene for tlyA, and its adjacent genetic loci were identified in all 17 C. lari isolates [n = 13 for UPTC; n = 4 for urease-negative (UN) C. lari]. Although sequence differences were demonstrated at approximately 20 loci within the 90 bp non-coding (NC) region, including the putative promoter structure candidates immediately upstream of the tlyA gene among the 18 isolates including C. lari RM2100, no sequence differences were identified within the NC region among the five UN C. lari isolates examined. A start codon ATG and a probable ribosome-binding site, AGGC(T)GG(A), for the tlyA gene were identified in all 18 isolates, including C. lari RM2100. The putative intrinsic ρ-independent transcriptional terminator structure candidate was also identified for the tlyA gene in both UPTC CF89-12 and C. lari RM2100. Additionally, the hemolysis assay was performed with some of the C. lari isolates. The tlyA gene nucleotide sequence data may possibly be useful for discrimination between UN C. lari and UPTC organisms, as well as for the differentiation among the four thermophilic Campylobacter species.

• MeSH
• Bacterial Proteins chemistry   genetics   MeSH
• Campylobacter lari chemistry   classification   genetics   isolation & purification   MeSH
• Phylogeny MeSH
• Hemolysin Proteins chemistry   genetics   MeSH
• Campylobacter Infections microbiology   veterinary   MeSH
• Chickens microbiology   MeSH
• Environmental Microbiology MeSH
• Bivalvia microbiology   MeSH
• Molecular Sequence Data MeSH
• Open Reading Frames MeSH
• Promoter Regions, Genetic MeSH
• Birds microbiology   MeSH
• Amino Acid Sequence MeSH
• Base Sequence MeSH
• Sequence Alignment MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Molecular cloning, nucleotide sequencing, and characterization of the flaA gene from additional isolates of urease-positive thermophilic Campylobacter (UPTC) were performed. These isolates were obtained from the natural environment in Northern Ireland (n = 9 from mussels) and in England (n = 1 from sea water). All isolates carried the shorter flaA gene, [open reading frames (ORFs), 1,461 to 1,503 base pairs], without any internal termination codons, and did not carry any flaA pseudogenes. The UPTC isolates were well discriminated by the neighbor joining (NJ) phylogenetic tree constructed based on the putative flaA genes ORFs nucleotide sequence information. In addition, the NJ tree constructed based on the flaA-short variable region sequence information discriminated the Campylobacter lari isolates with a similar degree of discrimination power.

• MeSH
• Campylobacter lari classification   genetics   isolation & purification   metabolism   MeSH
• Campylobacter classification   genetics   isolation & purification   metabolism   MeSH
• Flagellin chemistry   genetics   metabolism   MeSH
• Phylogeny MeSH
• Cloning, Molecular MeSH
• Bivalvia microbiology   MeSH
• Molecular Sequence Data MeSH
• Seawater microbiology   MeSH
• Open Reading Frames MeSH
• Polymerase Chain Reaction MeSH
• Recombinant Proteins chemistry   genetics   metabolism   MeSH
• Amino Acid Sequence MeSH
• Sequence Analysis, DNA methods   MeSH
• Sequence Alignment MeSH
• Urease metabolism   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• England MeSH
• Northern Ireland MeSH

We aimed to clarify if the thermophilic Campylobacter lari organisms including urease-negative (UN) C. lari and urease-positive thermophilic Campylobacter (UPTC) can be differentiated at the species and/or subspecies levels by employing the full-length flaA gene and flaA short variable region (SVR) nucleotide sequence information or not. Thermophilic Campylobacter isolates (n = 45) including UN C. lari (n = 17), UPTC (n = 18), and Campylobacter jejuni (n = 10) were well discriminated at the isolate level by the unweighted pair group method using arithmetic means analysis and neighbor joining procedures constructed based on the full-length flaA gene and flaA SVR nucleotide sequence information. Thus, these procedures may possibly be useful for epidemimological studies for C. lari and C. jejuni.

• MeSH
• Campylobacter jejuni classification   genetics   isolation & purification   MeSH
• Campylobacter lari classification   genetics   isolation & purification   MeSH
• DNA, Bacterial chemistry   genetics   MeSH
• Flagellin genetics   MeSH
• Genotype MeSH
• Humans MeSH
• Molecular Sequence Data MeSH
• Polymorphism, Genetic MeSH
• Sequence Analysis, DNA MeSH
• Cluster Analysis MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH