Generation of a Hybrid Sequence-Specific Single Stranded Deoxyribonuclease
Science 1987, 238, 1401-1403, 10.1126/science.3685986
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.
Metal: CaLigand type: UndefinedHost protein: Staphylococcal nucleaseAnchoring strategy: ---Optimization: ---Reaction: Hydrolytic cleavageMax TON: <1ee: ---PDB: ---Notes: Engineered sequence specificity