Mutagenicity testing on non-selectively cloned Chinese hamster ovary cells with the protein-mapping method

Chemical mutagenesis was studied on Chinese hamster ovary cells by protein mapping. Cell cultures were treated with methylnitrosourea and the cells were cloned in non-selective media. The proteins of single clones were separated by 2-dimensional electrophoresis and analysed for qualitative (electrophoretic mobility) and quantitative (staining intensity; presence/absence) changes in the protein patterns. The investigation included 26 clones derived from treated cells and 26 control clones. The total number of gene loci tested was calculated from the number of protein spots analysed: it amounted to about 33 800. The protein patterns revealed 2 alterations defined as qualitative variant proteins. No alteration of this type was found in the control group. The frequency of quantitative variant proteins was increased by more than 100% compared with the control group. Our results and theoretical considerations suggest that the cellular concentration of single proteins offers a sensitive parameter for mutagenicity testing.