Supporting data for "Dissecting the molecular determinants of the major multidrug efflux pump transporter MexB towards its antibiotic and physiological substrates: the extensively drug-resistant genotype PA154197 as a paradigm"

This dataset includes both the raw data and processed data resulting from analysis for my thesis titled with " Dissecting the molecular determinants of the major multidrug efflux pump transporter MexB towards its antibiotic and physiological substrates: the extensively drug-resistant genotype PA154197 as a paradigm ". PA154197 is a clinical strain isolated from a bloodstream infection at the Queen Mary Hospital in Hong Kong. To investigate the role of the RND efflux pump MexAB-OprM in cell physiology, I mainly conducted the measurement of virulence factors (PYO, proteolytic activities), in-vitro gene expression studies (RT-qPCR), in-vivo gene expression studies (promoter-lux reporter), the quantification of auto-inducer signals based on LC-MS/MS (3OC12-HSL, C4-HSL) or biosensor (AHQ). To study the MexB recognition determinants towards its substrates, a mexB mutant library was constructed using the native CRISPR-Cas system. Minimum inhibitory concentrations to different groups of antibiotics of these mutants were measured. The molecular docking analysis based on Autodock Vina was conducted to predict the possible ligand binding poses inside the distal binding pocket.

本数据集包含题为《解析主要多重耐药外排泵转运蛋白MexB针对其抗生素与生理底物的分子决定簇——以广泛耐药基因型PA154197为研究范式》的学位论文相关的原始数据与经分析处理后得到的数据。PA154197是从香港玛丽医院血流感染样本中分离得到的临床菌株。为探究RND型外排泵（RND efflux pump）MexAB-OprM在细胞生理过程中的作用，本研究主要开展了以下实验：毒力因子（PYO、蛋白水解活性）检测、体外基因表达研究（实时定量聚合酶链反应（real-time quantitative polymerase chain reaction, RT-qPCR））、体内基因表达研究（启动子-lux报告系统）、基于液相色谱-串联质谱（liquid chromatography-tandem mass spectrometry, LC-MS/MS）的自诱导信号分子（3OC12-HSL、C4-HSL）或生物传感器（AHQ）的定量分析。为明确MexB对底物的识别决定簇，本研究利用天然CRISPR-Cas系统（clustered regularly interspaced short palindromic repeats-associated protein system）构建了mexB突变体库，并测定了这些突变体对不同类别抗生素的最低抑菌浓度（minimum inhibitory concentration, MIC）。此外，本研究基于Autodock Vina开展分子对接分析，以预测远端结合口袋内配体的潜在结合构象。