Detection of p53 gene mutation: Analysis by single-strand conformation polymorphism and Cleavase fragment length polymorphism

We have generated a collection of clones containing single point mutations within the exon 5-9 hot spot regions of the p53 gene by using polymerase ch ain reaction (PCR) to amplify select regions of the gene from characterized cell lines. These clones were then used to address the sensitivity of muta tion detection using slab-gel single-strand conformation polymorphism (SSCP ) and Cleavase fragment length polymorphism (CFLP) assay systems. Both meth ods exhibited high sensitivities for the detection of mutations in cloned p 53 mutations in this study: 97% for CFLP and 94% for SSCP. In addition to r esulting in higher sensitivity of mutation detection, CFLP has the capabili ty to analyze longer fragments. In this study, CFLP identified five introni c mutations which were not investigated in the exon-specific SSCP assay. Th ese results agree with those found elsewhere and demonstrate that CFLP scan ning can have practical advantages when used for the identification of sequ ence alterations within the p53 gene.