Analysis of the p53 tumor suppressor gene by pyrosequencing

Tumor suppressor genes are implicated in cell cycle progression. Inactivati on of these genes predominantly occurs through mutations and/or allelic los s that involves both alleles. With inactivation by multiple mutations in a single gene, cloning of the amplified gene is necessary to determine whethe r the mutations reside on one ol both alleles. Using pyrosequencing, a rece ntly developed approach based on sequencing-by-synthesis, we studied geneti c variability in the p53 tumor suppressor gene and could quantify the ratio between the mutated and wild-type amplified fragments. Further-more, this sequencing technique also allows allelic determination of adjacent mutation s with no cloning of amplified fragments.