Transforming the Escherichia coli Trp repressor into a site-specific nuclease.

@article{Sutton1993TransformingTE,
  title={Transforming the Escherichia coli Trp repressor into a site-specific nuclease.},
  author={Christopher Sutton and Abhishek Mazumder and Celia C H Chen and David S. Sigman},
  journal={Biochemistry},
  year={1993},
  volume={32 16},
  pages={4225-30}
}
The Escherichia coli Trp repressor has been converted into an operator-specific nuclease by alkylating cysteine-49, inserted by site-directed mutagenesis, with 5-(iodoacetamido)-1,10-phenanthroline. In the presence of copper ion and thiol, high yields (> 50%) of double-stranded breaks of DNA are observed after a 20-min reaction. The high cleavage efficiency of this derivatized protein (Trp repressor E49C-OP) can be attributed to the proximity of cysteine-49 to the minor groove, the site of the… CONTINUE READING
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