高级检索
吴健, 穆瑞瑞, 任存邦, 王春阳, 李珏, 张卫东, 范清堂, 段广才. 志贺菌耐多药外排泵emrE基因作用分析[J]. 中国公共卫生, 2009, 25(6): 676-678. DOI: 10.11847/zgggws2009-25-06-18
引用本文: 吴健, 穆瑞瑞, 任存邦, 王春阳, 李珏, 张卫东, 范清堂, 段广才. 志贺菌耐多药外排泵emrE基因作用分析[J]. 中国公共卫生, 2009, 25(6): 676-678. DOI: 10.11847/zgggws2009-25-06-18
WU Jian, MU Rui-rui, REN Cun-bang, . Effect of genetically modified efflux pump emrE in isolated Shigella spp.[J]. Chinese Journal of Public Health, 2009, 25(6): 676-678. DOI: 10.11847/zgggws2009-25-06-18
Citation: WU Jian, MU Rui-rui, REN Cun-bang, . Effect of genetically modified efflux pump emrE in isolated Shigella spp.[J]. Chinese Journal of Public Health, 2009, 25(6): 676-678. DOI: 10.11847/zgggws2009-25-06-18

志贺菌耐多药外排泵emrE基因作用分析

Effect of genetically modified efflux pump emrE in isolated Shigella spp.

  • 摘要: 目的 对临床耐多药志贺菌株H24的1种小多重耐药性家族(SMR)外排泵emrE基因进行定向分子进化,研究基因突变增强emrE外排泵耐药性的潜力,调查志贺菌外排泵基因emrE基因垂直进化对临床耐药发展的影响。方法 通过用体外易错PCR(EP-PCR)方法对emrE基因进行随机突变实验,用微量稀释生长曲线法进行菌株的耐药性筛选,用荧光实时定量RT-PCR测定基因表达情况,应用DNA分析软件DNAStar进行生物信息学分析。结果 在31个突变子中筛选到了突变子ep2-emrE(DH5α),它的emrE基因有6个氨基酸突变,分别为Thr-28→Ala,Cys-39→Ser,Tyr-40→终止子,Ser-72→Arg,Leu-93→Phe,Ile-101→Phe。它不但增强四环素和红霉素的耐药能力,还产生新的氯霉素耐药能力。结论 尽管emrE基因较易获得新增耐药性,但在志贺菌对四环素、红霉素等抗生素耐药性发展中,emrE基因的进化突变没有被筛选,表明有其他能力更强的耐药基因在突变进化中起作用。

     

    Abstract: Objective To study the potential gene mutation increasing the drug resistance ability of emrE efflux pumpvia genetic modification of the emrE gene from clinical isolated Shigella spp.stra in H24,and to investigate the relationship between clinical drugresistance development of Shigella spp.and the vertical evolution of emrE efflux pump.Methods Gene emrE was mutated viaerror prone PCR.Drugres is tance of the strain was determined by microdilution growth curve method.The expression of the genes was measured by fluorescent quantitative RT-PCR and the bioinformatics analysis was conducted by DNAS tar.Results Among the 31 mutants,ep2-emrE showed the ability of increasing tetracyc line and ery thromyc in res is tance and the newly gained ability of chlorom ycetinres is tance indicating the mutational potent of the gene.Conclusion Although it is easy for gene emrE to achieve new drug resistant ability,the mutation and vertical evolution of the emrE gene is not selected in the development of tetracyc line and eryth romyc in resistance.So,there may be some more strong multid rug resis tance genes and mutation.

     

/

返回文章
返回