Sensitization of Campylobacter jejuni to fluoroquinolone and macrolide antibiotics by antisense inhibition of the CmeABC multidrug efflux transporter.
Sensitization of Campylobacter jejuni to fluoroquinolone and macrolide antibiotics by antisense inhibition of the CmeABC multidrug efflux transporter.
复制标题
通过反义抑制 CmeABC 多药外排转运蛋白使空肠弯曲杆菌对氟喹诺酮类和大环内酯类抗生素敏感。
DOI：
10.1093/jac/dkp067
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Zhang,Qijing
中科院分区：
文献类型：
--
作者：
Jeon,Byeonghwa;Zhang,Qijing
ObjectiveThe aim of this study was investigate the efficacy and efficiency of antisense mediated gene silencing by peptide nucleic acid（PNA）for specific inactivation of CmeABC multidrug efflux transporter in Campylobacter jejuni. Methods PNA was designed to bind to the meA transcript and to inhibit translation of CmeA，the periplasma component of RND-type CmeABC efflux transporter of C. jejuni.通过蛋白质印迹法测定CmeA产生的抑制。在CmeA特异性PNA（CmeA-PNA）的存在下，测定了临床上重要的抗生素（包括环丙沙星和红霉素）的MIC。与CmeA产量减少一致，CmeA-PNA使空肠弯曲菌菌株对环丙沙星和红霉素更敏感。在2 µM浓度下，CmeA-PNA导致空肠弯曲杆菌NCTC 11168中环丙沙星和红霉素的MIC分别降低8倍和4倍。CmeA-PNA还增加了对环丙沙星或红霉素耐药的空肠弯曲菌菌株对抗生素的敏感性。结论反义技术是一种抑制CmeABC多药外排转运蛋白功能的可行方法，可进一步用于控制耐药的空肠弯曲菌。
ObjectivesThe aim of this study was to investigate the feasibility and efficacy of antisense-mediated gene silencing by peptide nucleic acid (PNA) for specific inactivation of the CmeABC multidrug efflux transporter inCampylobacter jejuni.MethodsPNA was designed to bind to thecmeAtranscript and to inhibit the translation of CmeA, the periplasmic component of the RND-type CmeABC efflux transporter ofC.jejuni. Inhibition of CmeA production was determined by western blotting. MICs of clinically important antibiotics, including ciprofloxacin and erythromycin, were measured in the presence of the CmeA-specific PNA (CmeA-PNA).ResultsCmeA-PNA greatly reduced the expression level of CmeA. Consistent with the reduced CmeA production, CmeA-PNA renderedC.jejunistrains more susceptible to ciprofloxacin and erythromycin. At a concentration of 2 µM, CmeA-PNA resulted in 8- and 4-fold reductions in the MICs of ciprofloxacin and erythromycin, respectively, inC.jejuniNCTC 11168. CmeA-PNA also increased the susceptibility to the antibiotics inC.jejunistrains that were resistant to ciprofloxacin or erythromycin.ConclusionsAntisense technology is a feasible method to suppress the function of the CmeABC multidrug efflux transporter, which may be further exploited to control antibiotic-resistantCampylobacter.