DNA-binding proteins as site-specific nucleases

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

Research output: Contribution to journalArticle

22 Citations (Scopus)

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
Pages (from-to)335-342
Number of pages8
JournalMolecular Microbiology
Volume12
Issue number3
DOIs
StatePublished - May 1 1994
Externally publishedYes

Fingerprint

DNA-Binding Proteins
Site-Directed Mutagenesis
Cysteine
Carrier Proteins
Magnetic Resonance Spectroscopy
Binding Sites
X-Rays
Genome
Amino Acids
DNA
copper-1,10-phenanthroline
5-iodoacetamido-1,10-phenanthroline

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

DNA-binding proteins as site-specific nucleases. / Pan, Clark Q.; Landgraf, Ralf; Sigman, David S.

In: Molecular Microbiology, Vol. 12, No. 3, 01.05.1994, p. 335-342.

Research output: Contribution to journalArticle

Pan, Clark Q. ; Landgraf, Ralf ; Sigman, David S. / DNA-binding proteins as site-specific nucleases. In: Molecular Microbiology. 1994 ; Vol. 12, No. 3. pp. 335-342.
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