The alkaline single cell gel electrophoresis:: a new test for assessing DNA single strand breaks in Neurospora crassa

被引:6
作者
Bhanoori, M [1 ]
Venkateswerlu, G [1 ]
机构
[1] Osmania Univ, Dept Biochem, Hyderabad 500007, Andhra Pradesh, India
关键词
alkaline comet assay; Neurospora crassa; cadmium toxicity; hydrogen peroxide toxicity; fungal genotoxicity test;
D O I
10.1016/S0027-5107(98)00124-9
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The single cell gel electrophoresis (comet assay) is a potent technique in testing double and single strand breaks in DNA. In this paper, we present an application of alkaline comet assay to filamentous fungi for genotoxicological assessment of heavy metals for the first time. A wild strain of Neurospora crassa SLA 4200 was grown in presence of cadmium sulfate (CdSO4) (10 mu M and 100 mu M) for 12 h. Protoplasts from 12-h old mycelia were prepared by using Novozym 234 and DNA damage was evaluated by alkaline comet assay. Hydrogen peroxide (H2O2) (50 mu M and 100 mu M) was taken as an internal standard for DNA damage. Both CdSO4 and H2O2 induced significant single strand breaks in DNA. The results indicate that alkaline comet assay is a sensitive and rapid method for DNA damage analysis in filamentous fungal systems. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:29 / 34
页数:6
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