Generation of a hybrid sequence-specific single-stranded deoxyribonuclease

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Science  04 Dec 1987:
Vol. 238, Issue 4832, pp. 1401-1403
DOI: 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.

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