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利用单细胞凝胶电泳、流式细胞术和正向突变试验对中国仓鼠卵巢细胞的一个分离克隆中的诱变剂进行分析。

Analysis of mutagens with single cell gel electrophoresis, flow cytometry, and forward mutation assays in an isolated clone of Chinese hamster ovary cells.

作者信息

Wagner E D, Rayburn A L, Anderson D, Plewa M J

机构信息

College of Agricultural, Consumer and Environmental Sciences, Department of Crop Sciences, University of Illinois at Urbana-Champaign, Urbana, USA.

出版信息

Environ Mol Mutagen. 1998;32(4):360-8.

PMID:9882011
Abstract

We investigated the induction of DNA strand breaks in the single cell gel electrophoresis (SCGE or comet) assay with whole cell clastogenicity measured with flow cytometric analysis in cells from an isolated clone of the Chinese hamster ovary (CHO) AS52 cell line. Under identical treatment conditions the responses were compared with forward mutation at gpt using 2-acetoxyacetylaminofluorene (2AAAF), ultraviolet radiation (UV) and ethyl methanesulfonate (EMS). Cytotoxicity for each agent was evaluated in the SCGE and forward mutation assays. Forward mutation was 4-10-fold more sensitive than DNA strand breaks detected in the SCGE assay. For 2AAAF and EMS, the kinetics of the induction of genetic damage were similar for the three assays, although there were differences in sensitivity. With UV, the induction kinetics of gpt mutation differed from that expressed by SCGE and flow cytometric analysis. With the chemical mutagens 2AAAF and EMS, there was a high correlation between the SCGE assay and forward mutation and also between the SCGE assay and flow cytometry. There was no significant correlation between flow cytometry and forward mutation. With UV, only the SCGE assay and flow cytometry were correlated. Agent-specific variations in the intragenomic distribution of DNA damage for each mutagen was measured in the SCGE assay.

摘要

我们在中国仓鼠卵巢(CHO)AS52细胞系的一个分离克隆细胞中,通过单细胞凝胶电泳(SCGE或彗星试验)研究了DNA链断裂的诱导情况,并采用流式细胞术分析测量了全细胞致断裂性。在相同的处理条件下,将这些反应与使用2-乙酰氧基乙酰氨基芴(2AAAF)、紫外线辐射(UV)和甲磺酸乙酯(EMS)进行的gpt正向突变反应进行了比较。在SCGE和正向突变试验中评估了每种试剂的细胞毒性。正向突变比在SCGE试验中检测到的DNA链断裂敏感4至10倍。对于2AAAF和EMS,三种试验中遗传损伤诱导的动力学相似,尽管在敏感性上存在差异。对于UV,gpt突变的诱导动力学与SCGE和流式细胞术分析所显示的不同。对于化学诱变剂2AAAF和EMS,SCGE试验与正向突变之间以及SCGE试验与流式细胞术之间存在高度相关性。流式细胞术与正向突变之间没有显著相关性。对于UV,只有SCGE试验与流式细胞术相关。在SCGE试验中测量了每种诱变剂在基因组内DNA损伤分布的试剂特异性差异。

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