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[简单的光度法和荧光法检测不足以评估紫外线诱导的大肠杆菌中控制色氨酸合成的基因突变]

[Simple photometric and fluorometric assays are insufficient for assessing UV-induced mutagenesis in genes controlling tryptophan synthesis in Escherichia coli].

作者信息

Zherebtsov S V

机构信息

Institute of Cytology RAS, St. Petersburg.

出版信息

Tsitologiia. 2003;45(1):101-5.

Abstract

A new approach to detecting induced mutations was tested based on the assay of cell extracts and special growth media following cultivation of UV irradiated Escherichia coli cells. No correlation was found between the UV dose and the optical densities of cultural media or cell extracts prepared by Triton X-100 treatment. Blue fluorescence of concentrated cultural media varied with cell dose, according to a rather complex law, which differed substantially from the known dose-effect curves for induced mutations. Nevertheless, a certain extent of the brown staining of tryptophan containing medium could, presumably, serve as a quite sensitive indicator of the integral metabolic activity of bacteria grown in the medium. Besides, we observed that overnight lag phase cultures became gradually more transparent, when analysed in the spectrophotometer cuvette just after their dilution with fresh medium.

摘要

基于对紫外线照射后的大肠杆菌细胞进行培养后对细胞提取物和特殊生长培养基的检测,测试了一种检测诱导突变的新方法。未发现紫外线剂量与经Triton X - 100处理制备的培养基或细胞提取物的光密度之间存在相关性。浓缩培养基的蓝色荧光随细胞剂量而变化,遵循相当复杂的规律,这与已知的诱导突变剂量效应曲线有很大不同。然而,含有色氨酸的培养基的一定程度的棕色染色大概可以作为培养基中生长细菌整体代谢活性的相当敏感的指标。此外,我们观察到,过夜的延迟期培养物在用新鲜培养基稀释后立即在分光光度计比色皿中分析时,会逐渐变得更加透明。

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