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志贺菌成簇规律间隔短回文重复序列相关蛋白基因cas1和cas2研究 被引量:8

Clustered regularly interspaced short palindromic repeat associated protein genes cas1 and cas2 in Shigella
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摘要 目的:研究成簇规律间隔短回文重复序列(CRISPR)相关蛋白基因cas1和cas2在志贺菌中的分布,并分析cas1和cas2基因突变与细菌耐药的关系。方法采用PCR扩增196株志贺菌cas1和cas2基因。对3株志贺菌(Z23、2003135、2008113)的cas2、cas1(a)和cas1(b)基因进行测序,分析其突变与耐药之间的关系。结果196株志贺菌均检出CRISPR相关蛋白基因cas1和cas2。测序结果显示,cas2的一致性为96.44%,cas1(a)的一致性为97.61%,cas1(b)的一致性为96.97%。菌株2003135的cas1(b)基因有2个突变位点:3177129位点(C→G)及3177126位点(G→C),Z23、2008113对应位置没有突变。药敏结果显示菌株2003135的耐抗生素种类和耐抗生素数目均大于菌株Z23、2008113。结论志贺菌中CRISPR基因广泛存在。cas1(b)基因突变菌株与未突变株比较,耐药性增强。cas1(b)基因突变可能是引起耐药程度不同的原因之一。 Objective To detect the distribution of clustered regularly interspaced short palindromic repeat(CRISPR)associated protein genes cas1 and cas2 in Shigella and to understand the characteristics of CRISPR with relationship between CRISPR and related characteristics on drug resistance. Methods CRISPR associated protein genes cas1 and cas2 in Shigella were detected by PCR,with its products sequenced and compared.Results The CRISPR-associated protein genes cas1 and cas2 were found in all the 196 Shigella isolates which were isolated at different times and locations in China. Consistencies showed through related sequencing appepared as follows:cas2,cas1 (a) and cas1(b)were 96.44%,97.61%and 96.97%,respectively. There were two mutations including 3177129 site(C→G)and 3177126 site(G→C)of cas1(b)gene in 2003135 strain which were not found in the corresponding sites of Z23 and 2008113. Results showed that in terms of both susceptibility and antibiotic-resistance,strain 2003135 was stronger than Z23 and 2008113. Conclusion CRISPR system widely existed in Shigella,with the level of drug resistance in cas1(b) gene mutant strains higher than in wild strains. Cas1(b)gene mutation might be one of the reasons causing the different levels of resistance.
出处 《中华流行病学杂志》 CAS CSCD 北大核心 2014年第5期581-584,共4页 Chinese Journal of Epidemiology
关键词 志贺菌 成簇规律间隔短回文重复序列 耐药 Shigella Clustered regularly interspaced short palindromic repeat Resistance
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  • 1Pourcel C, Salvignol G, Vergnaud G. CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies [J]. Microbiology, 2005,151 (3) : 653-663.
  • 2Rowe-Magnus DA, Guerout AM, Mazel D. Bacterial resistance evolution by recruitment of super-integron gene cassettes [J]. Mol Microbiol, 2002,43 (6) : 1657-1669.
  • 3王颖芳,宋春花,段广才,郗园林.志贺菌敏感株诱导耐药与marOR基因突变关系[J].中国公共卫生,2012,28(3):352-353. 被引量:1
  • 4Clinical and Laboratory Standards Institute. Performance standards for antibiotic disk susceptibility tests. Eleventh edition [ S ]. CLSI, 2012.
  • 5宋春花,黄学勇,郗园林,张梅喜,段广才.志贺菌敏感株与基因转移耐多药株蛋白组学分析[J].中国公共卫生,2008,24(1):42-45. 被引量:9
  • 6Palmer KL,Gilmore MS. Multidrug-resistant enterococci lack CRISPR-cas[J]. MBio,2010,1 (4) :e00227-210.
  • 7Burley KM, Sedgley CM. CRISPR-Cas, a prokaryotic adaptive immune system, in endodontic, oral, and multidrug-resistant hospital-acquired Enterococcus faecalis [ J ]. J Endod, 2012,38 ( 11 ) : 1511-1515.
  • 8Marraffini LA, Sontheimer EJ. CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA [J]. Science, 2008,322(5909) : 1843-1845.

二级参考文献21

  • 1杨海燕,段广才,郗园林.主动外排系统acrAB在志贺菌中分布和表达[J].中国公共卫生,2005,21(6):685-687. 被引量:33
  • 2刘志远,许淑珍,马纪平.高耐庆大霉素粪肠球菌质粒接合转移试验的方法探讨[J].中华医院感染学杂志,2005,15(6):608-610. 被引量:10
  • 3PooleK.Efflux-mediatedmultiresistanceinGram-negativebacteria[J].ClinMicrobiolInfect,2004,10(1):12-26.
  • 4PerezA,CanleD,LatasaC,etal.Cloning,nucleotidesequencing,andanalysisoftheAcrAB-TolCeffluxpumpofEnterobactercloacaeanddeterminationofitsinvolvementinantibioticresistanceinaclinicalisolate[J].AntimicrobAgentsChemother,2007,51(9):3247-3253.
  • 5deCristobalRE,VincentPA,SalomonRA.MultidrugresistancepumpAcrAB-TolCisrequiredforhigh-level,Tet(A)-mediatedtetracyclineresistanceinEscherichiacoli[J].JournalofAntimicrobialChemotherapy,2006,58(5):31-36.
  • 6Adler-MoscaH,Luthy-HottensteinJ,MartinettiLG,etal.Developmentofresistancetoquinolonesinfivepatientswithcampylobacteriosistreatedwithnorfloxacinorciprofloxacin[J].EurJClinMicrobiolInfectDis,1991,10(11):953-957.
  • 7Marshak DR,Kadonaga JT,Burgess RR,et al.朱厚础等译.蛋白质纯化与鉴定实验指南[M].北京:科学出版社,2000,94
  • 8Berkelman T,Stenstedt T.2-D electrophoresis using immobilized pH gradients:principles & Methods[M].Amersham Pharmacia Biotech Inc Press,1998:14-25.
  • 9Neuhoff V,Arold N,Taube D,et al.Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250[J].Electrophoresis,1998,9(6):255-262.〔6〕 Shevchenko A,Wilm M,Vorm O,et al.Mass spectrometric sequencing of proteins Sil-stained polyacrylamide gels[J].Anal Chem,1996,68(5):850-858.
  • 10Shevchenko A,Wilm M,Vorm O,et al.Mass spectrometric sequencing of proteins Sil-stained polyacrylamide gels[J].Anal Chem,1996,68(5):850-858.

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  • 1徐婷,黄一灵,李耘,吕媛,夏文颖,顾兵,刘根焰,潘世扬.六家医院整合子介导宋内志贺菌多重耐药的形成和传播机制研究[J].中华临床感染病杂志,2013,6(3):138-143. 被引量:6
  • 2Pourcel C, Salvignol G, Vergnaud G. CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies [J]. Microbiology, 2005, 151 (Pt 3) : 653- 663.
  • 3Ishino Y, Shinagawa H, Makino K, et al. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product[J]. J Bacteriol, 1987,169(12) :5429-5433.
  • 4Jansen R, Embden JD, Gaastra W, et al. Identification of genes that are associated with DNA repeats in prokaryotes [J]. Mol Microbiol, 2002,43 (6) : 1565-1575.
  • 5Kunin V, Sorek R, Hugertholtz P. Evolutionary conservation of sequence and secondary structures in CKISPR repeats IJ]. Genome Biol, 2007,8 (4) : R61.
  • 6Kunin V, He S, Warnecke F, et al. A bacterial metapopulation adapts locally to phage predation despite global dispersal [J]. Genome Res, 2008,18 (2) : 293-297.
  • 7Lillestol RK, Redder P, Garrett RA, et al. A putative viral defence mechanism in archaeal cells[J]. Archaea, 2006,2 ( 1 ) :59-72.
  • 8Makarova KS,Haft DH, Barrangou R, et al. Evolution and classification of the CRISPR-Cas systems [J]. Nat Rev Microbiol, 2011,9 (6) : 467-477.
  • 9Deveau H, Garneau JE,Moineau S. CR/SPR/Cas system and its role in phage-bacteria interactions [J]. Annu Rev Microbiol, 2010,64: 475 -493.
  • 10Haft DH, Selengut J, Mongodin EF, et al. A guild of 45 CRISPR- associated (Cas) protein families and multiple CRISPR/Cas subtypes exist in prokaryotic genomes [J]. PLoS Comput Biol, 2005,1 (6) : e60.

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