ANALYSIS OF 7,12-DIMETHYLBENZEN[A]ANTHRACENE INDUCED MUTATIONS BY THE USE OF RESTRICTION SITE MUTATION TECHNIQUE IN THE RAT

Sinan SÜZEN*°, James M. PARRY**

*Ankara University, Faculty of Pharmacy, Department of Toxicology, 06100 Tandogan, Ankara, TURKEY. **School of Biological Sciences, University of Wales Swansea, UK.
°Corresponding Author

Summary :
The restriction site mutation (RSM) technique was applied as a methodology for the detection of base changes in restriction enzyme recognition sequences. Rats were exposed to 7,12-dimethylbenz[a]anthracene (DMBA) in two different doses. Mutations were analyzed in a variety of restriction enzyme recognition sequences of the rat p53 and ras genes. Mutations were detected in the BslI recognition sequences (CCTGTCCTGGG) in the high dose DMBA treated rats. DNA sequences analyses revealed that the induction of mutation was G?A transition at the tenth base of the CCTGTCCTGGG sequences. The developed method allows the detection of base pair substitutions, small insertions or deletions within the restriction enzyme recognition site in any gene for which the DNA sequence is known without selection of mutant phenotype.

Key words:
Restriction site mutation assay, DMBA, ras, p53.