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

Gareth Jenkins | Zendy

Open Access

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

Author(s) -

Gareth Jenkins

Publication year - 1999

Publication title -

mutagenesis

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.723

H-Index - 91

eISSN - 1464-3804

pISSN - 0267-8357

DOI - 10.1093/mutage/14.1.37

Subject(s) - restriction site , restriction enzyme , biology , restriction digest , genetics , restriction map , restriction fragment length polymorphism , gene , restriction fragment , mutagenesis , site directed mutagenesis , microbiology and biotechnology , intron , mutation , sequence analysis , polymerase chain reaction , nucleic acid sequence , mutant

This paper describes a rapid screening procedure for the detection of DNA sequence 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 wild-type sequence increases the sensitivity beyond that of PCR-RFLP analysis (10(-4)-10(-5) detectable here). In this paper we describe the rapid detection of induced in vivo mutations transforming the ApaI restriction site present in intron 6 of the mouse p53 gene to a unique AvaII site. The potential application of this method in other genes and organisms as a rapid screen for induced mutations is discussed.

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