Genetic heterogeneity of the dnaK gene locus including transcription terminator region (TTR) in Campylobacter lari.
Nucleotide sequences of approximately 3.1 kbp consisting of the full-length open reading frame (ORF) for grpE, a non-coding (NC) region and a putative ORF for the full-length dnaK gene (1860 bp) were identified from a urease-positive thermophilic Campylobacter (UPTC) CF89-12 isolate. Then, following the construction of a new degenerate polymerase chain reaction (PCR) primer pair for amplification of the dnaK structural gene, including the transcription terminator region of C. lari isolates, the dnaK region was amplified successfully, TA-cloned and sequenced in nine C. lari isolates. The dnaK gene sequences commenced with an ATG and terminated with a TAA in all 10 isolates, including CF89-12. In addition, the putative ORFs for the dnaK gene locus from seven UPTC isolates consisted of 1860 bases, and the four urease-negative (UN) C. lari isolates included C. lari RM2100 reference strain 1866. Interestingly, different probable ribosome binding sites and hypothetically intrinsic p-independent terminator structures were identified between the seven UPTC and four UN C. lari isolates, respectively. Moreover, it is interesting to note that 20 out of a total of 28 polymorphic sites occurred among amino acid sequences of the dnaK ORF from 11 C. lari isolates, identified to be alternatively UPTC-specific or UN C. lari-specific. In the neighbour-joining tree based on the nucleotide sequence information of the dnaK gene, C. lari forms two major distinct clusters consisting of UPTC and UN C. lari isolates, respectively, with UN C. lari being more closely related to other thermophilic campylobacters than to UPTC.
Shitara
M
M
Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Fuchinobe 1-17-71, Sagamihara 229-8501, Japan.
Tsuboi
Y
Y
Sekizuka
T
T
Tazumi
A
A
Moorei
J E
JE
Millar
B C
BC
Taneike
I
I
Matsuda
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002167:Campylobacter / Q000235:genetics; D044863:Campylobacter lari / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis*; D015723:Gene Library; D006360:Heat-Shock Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016415:Sequence Alignment; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D006360:Heat-Shock Proteins