首页> 美国卫生研究院文献>Antimicrobial Agents and Chemotherapy >Expression of Escherichia coli TehA gives resistance to antiseptics and disinfectants similar to that conferred by multidrug resistance efflux pumps.
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Expression of Escherichia coli TehA gives resistance to antiseptics and disinfectants similar to that conferred by multidrug resistance efflux pumps.

机译:大肠杆菌TehA的表达赋予了对抗菌剂和消毒剂的抗性类似于多药抗性外排泵所赋予的抗性。

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摘要

The genes tehAB located at 32.3 min on the Escherichia coli chromosome were initially identified by their ability to mediate resistance to potassium tellurite (128 micrograms of K2TeO3 per ml) when overexpressed with a high-copy-number plasmid. The genes encode an integral membrane protein (TehA) of 36 kDa with 10 putative transmembrane segments and a second protein (TehB) of 23 kDa. Overexpression of TehAB results in hypersensitivity to dequalinium CI and methyl viologen (paraquat). Expression of TehA alone gives similar hypersensitivity. Overexpression of TehA gave resistance to tetraphenylarsonium CI, ethidium bromide, crystal violet and proflavin. The efflux of ethidium, measured by fluorescence quenching, revealed that TehA transported ethidium at twice the control rate and 10% of the rate of the highly resistant efflux transporter Emr Eco. Addition of tellurite had no effect on ethidium transport. In addition to the ethidium transport assay, a proflavin fluorescence assay which was approximately 200-fold more sensitive was also used. TehA was also found to have proflavin efflux activity. The addition of TeO32- to the proflavin transport assay on TehA caused a 20% increase in transport rate. Both ethidium and proflavin transport were found to be energy dependent.
机译:最初通过在高拷贝数质粒中过表达时介导对亚碲酸钾抗性的能力(每毫升128微克K2TeO3)来鉴定位于大肠杆菌染色体32.3分钟的基因tehAB。这些基因编码一个36 kDa的整合膜蛋白(TehA),具有10个假定的跨膜片段和一个23 kDa的第二个蛋白(TehB)。 TehAB的过度表达导致对脱喹啉CI和甲基紫精(百草枯)过敏。单独表达TehA会产生类似的超敏反应。 TehA的过表达赋予了对四苯基ar CI,溴化乙锭,结晶紫和前黄素的抗性。通过荧光猝灭测量的乙炔外排量表明,TehA转运乙锭的速率是高耐药性外排转运蛋白Emr Eco的控制速率的两倍和10%。添加亚碲酸盐对乙转运没有影响。除了乙锭转运测定法外,还使用了灵敏度高约200倍的前黄素荧光测定法。还发现TehA具有黄素外排活性。在TehA的黄素转运分析中添加TeO32-导致转运速率提高20%。发现乙锭和黄酮转运都是能量依赖性的。

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