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A forward mutation assay using ampicillin-resistance in Escherichia coli designed for investigating the mutagenicity
D Bosworth1, C Crofton-Sleigh, S Venitt
1Section of Chemical Carcinogenesis, Institute of Cancer Research, Sutton, Surrey, UK.
Abstract:
The development of a bacterial mutation assay using forward mutation to ampicillin-resistance is described. It is a technically simple assay using Escherichia coli D494uvrB transformed with a multi-copy mutator plasmid pGW1700. Mutation is detected by an increase in the frequency of ampicillin-resistant colonies following treatment of bacteria with the test material during logarithmic growth. The determination of viable counts allows a correction factor to be applied to compensate for the effects of sample-induced growth enhancement or toxicity on the bacterial population. The assay has been tested with a range of reference mutagens. It is particularly sensitive to base-pair substitution mutagens, detecting these at doses equal to or less than those detected in the Salmonella/microsome assay or the SOS Chromotest. The assay also detects frameshift mutagens but with lower sensitivity than the Salmonella/microsome assay.