Restriction endonuclease BsoFI is sensitive to the 5'-methylation of deoxycytidines in its recognition sequence.
The isoschizomeric restriction endonucleases Fnu4HI and BsoFI cleave DNA at 5'-GCdecreasesNGC-3' sequences. Fnu4HI has been shown to be inhibited by 5'-CG-3'methylation in the sequences 5'-GmCGGC-3' or 5'-GCGGmCG-3'. We have now investigated the methylation sensitivity of BsoFI by testing its activity on plasmid DNA 5'-CG-3' methylated with the M.SssI DNA methyltransferase or on synthetic (CGG)n repetitive oligodeoxyribonucleotides which have been partly or completely C methylated. The data demonstrate that BsoFI cannot cleave at its recognition sequence when it is completely 5'-CG-3' methylated. These enzymes have proven to be useful in analyses of the methylation status in (CGG)n repeats of the human genome.
Deissler
H
H
Institut für Genetik, Universität zu Köln, Germany.
Genç
B
B
Doerfler
W
W
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D003841:Deoxycytidine / Q000031:analogs & derivatives; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D008745:Methylation; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D013379:Substrate Specificity
D009838:Oligodeoxyribonucleotides; D003841:Deoxycytidine; D004247:DNA; C016569:5-methyldeoxycytidine; C069758:endodeoxyribonuclease Fnu4HI; D015252:Deoxyribonucleases, Type II Site-Specific