Mutagen testing using TRP+ reversion in Escherichia coli

Escherichia coli strain WP2 and its repair-deficient derivatives are suitable strains for mutagen screening. In these strains, agents which cause base substitution mutations can be shown to increase the frequency of Trp+ revertants. In addition, agents causing many types of DNA damage can be detected through increased killing of the repair deficient derivatives. Four ways of performing tests are described: (a) Spot tests in which a small amount of the agent under test is placed directly on a selective agar plate. Trp+ revertants are counted and increased sensitivity of repair-deficient strains determined from the size of the zone of inhibition of cell growth. (b) Treat and plate tests, where a strain is treated with the agent under test and subsequently plated to determine survival or frequency of Trp+ revertants. (c) A simplified fluctuation test which shows exceptional sensitivity in measuring mutation with low levels of mutagens. (d) Use of a liver microsomal fraction in conjunction with treat and plate tests to detect metabolically activated mutagens. The merits and defects of these systems are discussed. Common pitfalls in evaluating tests and procedures for avoiding them are described.