The structure of the rare-cutting restriction endonuclease NotI, which recognizes the 8 basepair target 5′-GCGGCCGC-3′, has been solved with and without bound DNA. Because of its specificity (recognizing a site that occurs once per 65kb), NotI is used

Restriction endonucleases (REases) with 8-base specificity are rare specimens in nature. NotI from Nocardia otitidis-caviarum (recognition sequence 5′-GCGGCCGC-3′) has been cloned, thus allowing for mutagenesis and screening for enzymes with altered

The restriction endonuclease (ENase) NotI is blocked by methylation within its recognition sequence at 5'-GCGGCmCGC-3'. This sensitivity to methylation can be used to enhance the specificity of NotI in vivo and in vitro. Modification by M.FnuDII or