title: Molecular identification and characterization of clustered regularly interspaced short palindromic repeat (CRISPR) gene cluster in Taylorella equigenitalis.
abstract: Clustered regularly interspaced short palindromic repeats (CRISPRs), of approximately 10,000 base pairs (bp) in length, were shown to occur in the Japanese Taylorella equigenitalis strain, EQ59. The locus was composed of the putative CRISPRs-associated with 5 (cas5), RAMP csd1, csd2, recB, cas1, a leader region, 13 CRISPR consensus sequence repeats (each 32 bp; 5'-TCAGCCACGTTCGCGTGGCTGTGTGTTTAAAG-3'). These were in turn separated by 12 non repetitive unique spacer regions of similar length. In addition, a leader region, a transposase/IS protein, a leader region, and cas3 were also seen. All seven putative open reading frames carry their ribosome binding sites. Promoter consensus sequences at the -35 and -10 regions and putative intrinsic ρ-independent transcription terminator regions also occurred. A possible long overlap of 170 bp in length occurred between the recB and cas1 loci. Positive reverse transcription PCR signals of cas5, RAMP csd1, csd2-recB/cas1, and cas3 were generated. A putative secondary structure of the CRISPR consensus repeats was constructed. Following this, CRISPR results of the T. equigenitalis EQ59 isolate were subsequently compared with those from the Taylorella asinigenitalis MCE3 isolate.
authors.affiliation: Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Fuchinobe 1-17-71, Chuo-ku, Sagamihara, 252-5201, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Hara
authors.forename: Kyohei
authors.initials: K
authors.lastname: Hayashi
authors.forename: Takuya
authors.initials: T
authors.lastname: Nakajima
authors.forename: Shizuko
authors.initials: S
authors.lastname: Kagawa
authors.forename: Akihiro
authors.initials: A
authors.lastname: Tazumi
authors.forename: John E
authors.initials: JE
authors.lastname: Moore
authors.forename: Motoo
authors.initials: M
authors.lastname: Matsuda
mesh_terms: D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D020869:Gene Expression Profiling; D007564:Japan; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016366:Open Reading Frames; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D020625:Taylorella equigenitalis / Q000235:genetics* / Q000302:isolation & purification; D061806:Transcription Termination, Genetic
chemical_list: D004269:DNA, Bacterial