College of Animal Science and Technology, Guangxi University, Nanning 530005, People's Republic of China.
Guangxi Nongken Yongxin Animal Husbandry Group Xijiang Co. LTD, Guigang 537000, People's Republic of China.
J Biochem. 2021 Feb 6;169(1):101-108. doi: 10.1093/jb/mvaa100.
The two-component system BaeSR participates in antibiotics resistance of Escherichia coli. To know whether the outer membrane proteins involve in the antibiotics resistance mediated by BaeSR, deletion of acrB was constructed and the recombined plasmid p-baeR was introduced into E. coli K12 and K12△acrB. Minimum inhibitory concentrations (MICs) of antibacterial agents were determined by 2-fold broth micro-dilution method. Gene expressions related with major outer membrane proteins and multidrug efflux pump-related genes were determined by real-time quantitative reverse transcription polymerase chain reaction. The results revealed that the MICs of K12ΔacrB to the tested drugs except for gentamycin and amikacin decreased 2- to 16.75-folds compared with those of K12. When BaeR was overexpressed, the MICs of K12ΔacrB/p-baeR to ceftiofur and cefotaxime increased 2.5- and 2-fold, respectively, compared with their corresponding that of K12△acrB. At the same time, the expression levels of ompC, ompF, ompW, ompA and ompX showed significant reduction in K12ΔacrB/p-baeR as compared with K12△acrB. Moreover, the expression levels of ompR, marA, rob and tolC also significantly 'decreased' in K12ΔacrB/p-baeR. These findings indicated that BaeR overproduction can decrease cephalosporins susceptibility in acrB-free E. coli by decreasing the expression level of outer membrane proteins.
双组份系统 BaeSR 参与大肠杆菌的抗生素耐药性。为了了解外膜蛋白是否参与 BaeSR 介导的抗生素耐药性,构建了 acrB 的缺失突变体,并将重组质粒 p-baeR 导入大肠杆菌 K12 和 K12△acrB。采用二倍肉汤微量稀释法测定抗菌药物的最小抑菌浓度(MIC)。通过实时定量逆转录聚合酶链反应测定与主要外膜蛋白和多药外排泵相关基因的基因表达。结果显示,与 K12 相比,K12△acrB 对除庆大霉素和阿米卡星以外的测试药物的 MIC 值降低了 2 至 16.75 倍。当 BaeR 过表达时,K12△acrB/p-baeR 对头孢噻呋和头孢噻肟的 MIC 值分别增加了 2.5 倍和 2 倍,而 K12△acrB 的 MIC 值则相应增加。同时,与 K12△acrB 相比,K12△acrB/p-baeR 中 ompC、ompF、ompW、ompA 和 ompX 的表达水平显著降低。此外,在 K12△acrB/p-baeR 中,ompR、marA、rob 和 tolC 的表达水平也显著“降低”。这些发现表明,BaeR 的过表达可以通过降低外膜蛋白的表达水平降低无 acrB 大肠杆菌对头孢菌素的敏感性。