Site-specific Mutagenesis Approach

This powerful experimental approach involves a strategy in which a shuttle plasmid vector containing a single defined DNA adduct or lesion is allowed to replicate in mammalian cells or bacteria. The precise position of mutations arising from a given lesion then is established by DNA sequence analysis. A double strand shuttle vector that replicates in bacteria and yeast or human cells provides mechanistic information for site specific studies (Levine et al.)