Yuan Pengfei, Dong Junqing, Zhao Weibin, Zhuo Min, Li Shuang, Huang Shaobin, Li Jianjun
School of Biology and Biological Engineering, South China University of Technology, Higher Education Mega Center Guangzhou 510006 China
School of Environment and Energy, South China University of Technology, Higher Education Mega Center Guangzhou 510006 China.
RSC Adv. 2019 Nov 4;9(61):35662-35670. doi: 10.1039/c9ra06202e. eCollection 2019 Oct 31.
A new bacterial genotoxicity detection strain was constructed, in which the cell lysis gene of from a lambda phage was controlled by a new designed SOS responsive element, designated as BL21/pUC-PST. The biosensor responded only after 0.5 h contact with mutagens and the changes in cell culture turbidity could be easily differentiated with the naked eyes from the control sample. This SOS/ system presented a dose-dependent manner to five model DNA-damaging agents with an improved detection sensitivity. The limits of detection (LODs) were 0.026 μM for mitomycin C, 320.4 μM for azinphos-methyl, 34.4 μM for methyl methanesulfonate, 4.6 μM for dithianone and 6.0 μM for dichlofluanid, which were much lower than previously reported. By performing binary and ternary mixture experiments, the toxic equivalency concept was validated in the SOS/ system by comparison with bioanalytical equivalent concentrations (BEQ) and overall toxic equivalent concentration (TEQ) using Cr(vi) as the reference compound. Pearson analysis indicated that a strong correlation existed between the TEQ and BEQ values. Thus the TEQ could be presented as the Cr(vi) equivalent concentration from its dose-effect lysis profiles for the environmental sample. The proposed genotoxicity reporter strain allows for easier qualitative characterization and quantitative interpretation of the TEQ values using Cr(vi) as the reference for environmental water samples.
构建了一种新的细菌遗传毒性检测菌株,其中来自λ噬菌体的细胞裂解基因由新设计的SOS响应元件控制,命名为BL21/pUC-PST。该生物传感器仅在与诱变剂接触0.5小时后才有反应,并且细胞培养物浊度的变化可以很容易地用肉眼与对照样品区分开来。这种SOS/系统对五种模型DNA损伤剂呈现剂量依赖性方式,检测灵敏度有所提高。丝裂霉素C的检测限(LOD)为0.026 μM,谷硫磷为320.4 μM,甲基磺酸甲酯为34.4 μM,二噻农为4.6 μM,百菌清为6.0 μM,均远低于先前报道的值。通过进行二元和三元混合物实验,以Cr(Ⅵ)为参考化合物,通过与生物分析等效浓度(BEQ)和总毒性等效浓度(TEQ)比较,在SOS/系统中验证了毒性当量概念。Pearson分析表明TEQ和BEQ值之间存在强相关性。因此,对于环境样品,TEQ可以从其剂量效应裂解曲线表示为Cr(Ⅵ)等效浓度。所提出的遗传毒性报告菌株允许使用Cr(Ⅵ)作为环境水样的参考,更轻松地对TEQ值进行定性表征和定量解释。