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脲阳性嗜热弯曲杆菌(UPTC)中串联重复短回文序列(CRISPRs)的分子鉴定与特征分析。

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

机构信息

Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Fuchinobe 1-17-71, Chuo-ku, Sagamihara, 252-5201, Japan.

出版信息

World J Microbiol Biotechnol. 2012 Feb;28(2):713-20. doi: 10.1007/s11274-011-0867-3. Epub 2011 Aug 28.

Abstract

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.

摘要

新型簇状规律间隔短回文重复序列(CRISPRs)基因座[7500 个碱基对(bp)]存在于产脲阳性嗜热弯曲菌(UPTC)日本分离株 CF89-12 中。7500bp 基因座包含 5'-甲基氨甲酰基-2-硫代尿嘧啶甲基转移酶基因、假定 CRISPR 相关(p-Cas)、假定开放阅读框、Cas1 和 Cas2、前导序列区(146bp)、12 个 CRISPR 共有序列重复(每个 36bp),由类似长度(26-31bp)的非重复独特间隔区隔开,以及磷脂酰甘油磷酸酶 A 基因。当 UPTC CF89-12 和五株空肠弯曲菌分离株的 CRISPRs 基因座相互比较时,这六株分离株在基因座内均含有 p-Cas、Cas1 和 Cas2。6 株分离株中出现 4 到 12 个 CRISPR 共有序列重复,由非重复独特间隔区隔开,这些重复的核苷酸序列彼此之间具有约 92-100%的相似性。然而,这些分离株之间的独特间隔区没有序列相似性。检测到 CRISPRs 和 Cas 的假定 σ(70)转录启动子和假设 ρ 非依赖性终止子结构。未在 UPTC 细胞中证实 p-Cas、Cas1 和 Cas2 的体内转录。

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