In vitro sodium bisulfite mutagenesis of restriction endonuclease recognition sites.
Sodium bisulfite treatment of single-stranded DNA deaminates exposed cytosine residues to form uracil, resulting in cytosine-to-thymidine transition mutations following DNA replication. We have used this reaction in vitro to destroy the recognition sequences for the restriction endonucleases HindIII and XmaI in the aminoglycoside 3'-phosphotransferase I coding region of plasmid pUC4K. This procedure should be applicable to the mutation of any recognition sequence of restriction endonucleases which generate cytosine-containing single-stranded ends. The possibility of mutagenesis of restriction sites to generate stop codons in coding regions is discussed.
D001483:Base Sequence; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004269:DNA, Bacterial / Q000187:drug effects*; D004277:DNA, Single-Stranded / Q000187:drug effects*; D004926:Escherichia coli / Q000235:genetics; D005815:Genetic Code; D019868:Kanamycin Kinase; D009154:Mutation; D010770:Phosphotransferases / Q000235:genetics*; D010957:Plasmids; D013447:Sulfites / Q000494:pharmacology*; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D013447:Sulfites; D010770:Phosphotransferases; D019868:Kanamycin Kinase; D004262:DNA Restriction Enzymes; C009279:sodium bisulfite