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O157:H7型大肠埃希菌CRISPR结构及耐药基因和毒力基因分布分析

The CRISPR structures and the distribution of virulence genes and resistance genes in Escherichia coli O157:H7
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摘要 目的了解O157:H7型大肠埃希菌中成簇的规律间隔短回文重复序列(Clustered Regularly Interspaced ShortPalindromic Repeats,CRISPR)结构特征以及耐药基因和毒力基因分布情况。 方法利用多序列比对、BLAST、RNA二级结构预测等多种生物信息学方法对20株O157:H7型大肠埃希菌的全基因组序列进行分析。 结果从GenBank数据库获得2000-2014年20株O157:H7型大肠埃希菌全基因组序列,所有基因组中共存在3个CRISPR位点且每个CRISPR位点的基因序列具有高度一致性(均超过99.7%);CRISPR2被一段序列分隔为CRISPR2a和CRISPR2b;CRISPR1中存在3个不同的间隔序列,CRISPR2也有3个不同的间隔序列,CRISPR3有1个间隔序列;CRISPR1和CRISPR3的重复序列均可形成茎环结构,CRISPR2和CRISPR1的重复序列具有较高的一致性。共检测7个耐药基因,所有菌株ampc均阳性,4株菌株tetB和sul1阳性,其它4个基因均未检出;共检测6个毒力基因,有10株细菌stx1阳性,其他5个毒力基因20株细菌均阳性。 结论CRISPR在O157:H7型大肠埃希菌中广泛分布且结构稳定,毒力基因分布广,耐药性问题不可忽视。 Objective To ascertain the structure of clustered regularly interspaced short palindromic repeats (CRISPR)and the distribution of virulence genes and resistance genes of Escherichia coli O157:H7. Methods The genomic sequences of a total of 20strains of E.coli O157:H7were analyzed via multiple sequence alignment,BLAST,and RNA secondary structure prediction. Results The genomic sequences of 20strains of E.coli O157:H7were downloaded from the National Center for Biotechnology information(NCBI)database from 2000to 2014.All genomes contained 3 CRISPR loci,and the gene sequences of each CRISPR locus were highly consistent(all over 99.7%).CRISPR2was divided into two types based on fragments,designated CRISPR2aand CRISPR2b.There were three different spacers in CRISPR1and CRISPR2but only one spacer in CRISPR3.The repeats in CRISPR1and CRISPR3form a stem-loop structure, and the repeats in CRISPR2and CRISPR1are highly consistent.Of the 7drug resistance genes detected,ampc was found in all strains,tetBand sul1were found in 4strains,and the other 4genes were not detected in any strain.Of the 6 virulence genes tested,stx1was found in 10strains,and the other 5genes were detected in all 20strains. Conclusion CRISPR are widely distributed in E.coli O157:H7,with a stable structure and wide distribution of virulence genes.The problem of drug resistance cannot be ignored.
作者 王鹏飞 王颖芳 段广才 郗园林 徐亚珂 陈帅印 詹煜慧 WANG Peng-fei;WANG Ying-fang;DUAN Guang-cai;XI Yuan-lin;XU Ya-ke;CHEN Shuaiyin;ZHAN Yu-hui(Henan University of Science and Technology Hospital,Luoyang,China 471023;Department of Public Health,College of Medicine,Henan University of Science and Technology;Department of Epidemiology,College of Public Health,Zhengzhou University;College of Medicine,Henan University of Science and Technology)
出处 《中国病原生物学杂志》 CSCD 北大核心 2019年第7期761-764,772共5页 Journal of Pathogen Biology
基金 国家科技重大专项(No.2013ZX10004607,2018ZX10301407)。
关键词 O157:H7型大肠埃希菌 CRISPR 毒力基因 耐药基因 Escherichia coli O157:H7 CRISPR resistance gene virulence gene
作者简介 王鹏飞(1989-),男,河南洛阳人,硕士研究生,医师,主要研究方向为分子流行病学。E-mail:603971586@qq.com;通讯作者:段广才,E-mail:gcduan@zzu.edu.cn.
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