DNA‐binding proteins as site‐specific nucleases

Clark Q. Pan, Rail Landgraf, David S. Sigman

Research output: Contribution to journalReview article

22 Scopus citations

Abstract

DNA-binding proteins can be converted into site-specific nucleases by linking them to the chemical nuclease 1,10-phenanthroline-copper. This can be readily accomplished by converting a minor groove-proximal amino acid to a cysteine residue using site-directed mutagenesis and then chemically modifying the sulphydryl group with 5-iodoacetamido-1,10-phenanthroline- copper. These chimeric scission reagents can be used as rare cutters to analyse chromosomal DNA, to test predictions based on high-resolution nuclear magnetic resonance and X-ray crystal structures, and to locate binding sites of proteins within genomes.

Original languageEnglish (US)
Pages (from-to)335-342
Number of pages8
JournalMolecular Microbiology
Volume12
Issue number3
DOIs
StatePublished - May 1994
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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