Molecular identification and characterization of clustered regularly interspaced short palindromic repeats (CRISPRs) in a urease-positive thermophilic Campylobacter sp. (UPTC)

• Published in: World journal of microbiology & biotechnology 2012-02, Vol.28 (2), p.713-720
• Main Authors: Tasaki, E., Hirayama, J., Tazumi, A., Hayashi, K., Hara, Y., Ueno, H., Moore, J. E., Millar, B. C., Matsuda, M.
• Format: Article
• Language: English
• Subjects: Analysis; Applied Microbiology; Bacteria; Biochemistry; Biomedical and Life Sciences; Biotechnology; Campylobacter; Campylobacter - enzymology; Campylobacter - genetics; Chemical synthesis; CRISPR; Environmental Engineering/Biotechnology; Fetuses; Gene expression; Gene loci; Genes, Bacterial - genetics; Genetic testing; Genomes; Inverted Repeat Sequences - genetics; Laboratories; Life Sciences; Microbiology; Organisms; Original Paper; Public health; Repetitive Sequences, Nucleic Acid - genetics; Reverse Transcriptase Polymerase Chain Reaction; Studies; Urease - metabolism
• ISSN: 0959-3993; 1573-0972
• DOI: 10.1007/s11274-011-0867-3

Novel clustered regularly-interspaced short palindromic repeats (CRISPRs) locus [7,500 base pairs (bp) in length] occurred in the urease-positive thermophilic Campylobacter (UPTC) Japanese isolate, CF89-12. The 7,500 bp gene loci consisted of the 5′-methylaminomethyl-2-thiouridylate methyltransferase gene, putative (P) CRISPR associated (p- Cas ), putative open reading frames, Cas1 and Cas2 , leader sequence region (146 bp), 12 CRISPRs consensus sequence repeats (each 36 bp) separated by a non-repetitive unique spacer region of similar length (26–31 bp) and the phosphatidyl glycerophosphatase A gene. When the CRISPRs loci in the UPTC CF89-12 and five C. jejuni isolates were compared with one another, these six isolates contained p- Cas , Cas1 and Cas2 within the loci. Four to 12 CRISPRs consensus sequence repeats separated by a non-repetitive unique spacer region occurred in six isolates and the nucleotide sequences of those repeats gave approximately 92–100% similarity with each other. However, no sequence similarity occurred in the unique spacer regions among these isolates. The putative σ 70 transcriptional promoter and the hypothetical ρ-independent terminator structures for the CRISPRs and Cas were detected. No in vivo transcription of p- Cas , Cas1 and Cas2 was confirmed in the UPTC cells.