Generation of a hybrid sequence-specific single-stranded deoxyribonuclease.
The relatively nonspecific single-stranded deoxyribonuclease, staphylococcal nuclease, was selectively fused to an oligonucleotide binding site of defined sequence to generate a hybrid enzyme. A cysteine was substituted for Lys116 in the enzyme by oligonucleotide-directed mutagenesis and coupled to an oligonucleotide that contained a 3'-thiol. The resulting hybrid enzyme cleaved single-stranded DNA at sites adjacent to the oligonucleotide binding site.
Corey
D R
DR
Department of Chemistry, University of California, Berkeley 94720.
Schultz
P G
PG
D000595:Amino Acid Sequence; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000378:metabolism; D006868:Hydrolysis; D008836:Micrococcal Nuclease / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity
D004277:DNA, Single-Stranded; D008836:Micrococcal Nuclease