Inverse restriction site mutation (iRSM) analysis. Mutation detection involving the formation of restriction enzyme sites in target genes

This paper describes a rapid screening procedure for the detection of DNA s equence changes resulting in the creation of new restriction enzyme sites. The basic methodology involves the identification of the conversion of one restriction site into another by mutagenesis. The selective removal of the wild-type sequences by digestion with a restriction enzyme acting on the wi ld-type sequence increases the sensitivity beyond that of PCR-RFLP analysis (10(-4)-10(-5) detectable here). In this paper we describe the rapid detec tion of induced in vivo mutations transforming the ApaI restriction site pr esent in intron 6 of the mouse p53 gene to a unique AvaII site. The potenti al application of this method in other genes and organisms as a rapid scree n for induced mutations is discussed.