Additionally, false positives (i.e. non-carcinogens detected as mutagens) do occur GSK126 in vitro in the Ames test. There are a small number of compounds that are Ames positive mutagens due to their bacterium-specific metabolism e.g. sodium azide and some nitro-group containing compounds (Prival, 1983). The strains of Salmonella typhimurium used in the Ames test contain different mutations in various histidine synthesis genes ( Table 1). The mutations carried by the specific strains prevent the bacteria from growing in media without histidine. However, if the test chemical mutates the defective mutation

back to functional status (revert initial mutation), the bacteria will acquire the ability to grow in histidine-free media and form colonies. These colonies are thus known as revertants ( Ames et al., 1975). All strains except TA102 are missing the uvrB DNA repair gene, thus removing the main error-free DNA excision repair pathway, compared to wild-type cells. This will amplify the mutations as DNA repair, in the absence of excision repair, occurs by error-prone

pathways. TA102 bacteria strain maintains the excision repair system to be able to detect DNA cross-linking agents such as mitomycin C. Otherwise compounds with DNA cross-link mechanism of action will not be detected, as unrepaired cross-links are lethal to the cell. In addition, all strains have the mutation known as deep rough or rfa genotype. This is an alteration of the phenotype, where the polysaccharide capsule surrounding the cell is no longer www.selleckchem.com/products/atezolizumab.html present. Therefore, larger compounds are able to enter through the cell membrane reaching the bacterial DNA. Various strains possess the plasmid pKM101 which contains the operon muc. Enzymes encoded by this operon allow the damaged DNA to continue its synthesis. The effect of Ribose-5-phosphate isomerase this operon is to amplify the translation of DNA damage to mutations. The plasmid also contains a gene coding for resistance to the antibiotic ampicillin. This ampicillin-resistant property

permits the selection of mutants containing the plasmid. Alternatively, some Escherichia coli strains can be used to screen for mutagens. These strains have base change mutations in one of the tryptophan synthesis operon genes (trpE) instead of the histidine operon genes. Strains with and without the uvrA mutation are available as are strains with and without the plasmid pKM101. E. coli WP2 strains are equivalent to TA102 in terms of types of mutagen detected (including oxidative mutagens). However, if a cross-linking effect is to be detected, then the E. coli strain must have an intact excision repair system. The rfa mutation is not required as E. coli cells are naturally permeable to larger molecules. Each strain of bacteria used in the Ames test detects a different spectrum of mutagens.