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The application of a DNA damage assay for detecting environmental genotoxic pollutants.

机译：DNA损伤测定法在检测环境遗传毒性污染物中的应用。

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摘要

An in vitro assay system for detecting cellular DNA damage in FKM (fathead minnow) exposed to genotoxicants, HN2 (nitrogen mustard) and MNNG (N-methyl,N{dollar}spprime{dollar}-nitro-N-nitrosoguanidine), is described. DNA from the exposed FHM was examined for cellular DNA strand breakage by agarose gel electrophoresis. FHM were exposed to a series of nitrogen mustard concentrations (0, 10, 20, 30, 50, 100, 200, 500 mg/L) dissolved in MHW (moderately hard water) for 2 hr. The DNA was isolated, purified and checked with the spectrophotometer so that the OD260/OD280 ratio was not less than 1.6. DNA samples were then separated by 0.7% agarose gel electrophoresis for 2 hr at a constant voltage of 50. The gel was then photographed with a IS-1000 Digital Imaging System. The cellular DNA bands were identified and quantified with the ImageQuaNT{dollar}rmsp{lcub}TM{rcub}{dollar} software. The control group's DNA content, set at 100%, was compared to the DNA derived from various HN2 exposures. A dose-dependent response curve based on DNA content was observed from which an EC50 value was derived. Similarly, the MNNG exposure study performed at comparable concentrations yielded a dose-dependent curve based on DNA content. Samples from a wastewater treatment plant and a metal planting company were utilized to demonstrate for the DNA damage assay in fathead minnows. This assay proved to be a more sensitive indicator for evaluating aquatic pollutant when compared to the standard acute toxicity routinely employed. Based on these laboratory data, it appears that this in vitro assay could have applicability in screening for aquatic genotoxic pollutant in the environment. The relationships between acute (LC50) and chronic endpoints of aquatic organisms and this in vitro assay were investigated and discussed.
机译：描述了一种体外测定系统，该系统用于检测暴露于遗传毒物HN2（氮芥）和MNNG（N-甲基，N {美元} spprime {dollar}-硝基-N-亚硝基胍）的FKM（无头min鱼）中的细胞DNA损伤。通过琼脂糖凝胶电泳检查来自暴露的FHM的DNA的细胞DNA链断裂。将FHM暴露于溶解在MHW（中度硬水）中的一系列氮芥末浓度（0、10、20、30、50、100、200、500 mg / L）2小时。分离DNA，纯化并用分光光度计检查，以使OD260 / OD280之比不小于1.6。然后将DNA样品通过0.7％琼脂糖凝胶电泳在50恒定电压下分离2小时。然后用IS-1000 Digital Imaging System对凝胶照相。使用ImageQuaNT {dollar} rmsp {lcub} TM {rcub} {dollar}软件对细胞DNA带进行鉴定和定量。将对照组的DNA含量（设置为100％）与来自各种HN2暴露的DNA进行比较。观察到基于DNA含量的剂量依赖性反应曲线，从中得出EC50值。同样，以相当的浓度进行的MNNG暴露研究得出了基于DNA含量的剂量依赖性曲线。利用废水处理厂和一家金属种植公司的样品证明了黑头min鱼的DNA损伤测定。与常规采用的标准急性毒性相比，该测定方法被证明是评估水生污染物的更灵敏指标。基于这些实验室数据，看来该体外测定法可用于筛选环境中的水生遗传毒性污染物。研究和讨论了水生生物急性（LC50）和慢性终点之间的关系以及该体外测定。

著录项

  • 作者

    Lan, Cheng-Hang.;

  • 作者单位

    University of Michigan.;

  • 授予单位 University of Michigan.;
  • 学科 Environmental Sciences.
  • 学位 Ph.D.
  • 年度 1990
  • 页码 130 p.
  • 总页数 130
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 环境科学基础理论;
  • 关键词

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