Site-directed mutagenesis in the catalytic center of the restriction endonuclease EcoRI.

@article{Grabowski1995SitedirectedMI,
  title={Site-directed mutagenesis in the catalytic center of the restriction endonuclease EcoRI.},
  author={Grzegorz Grabowski and Albert Jeltsch and Heiner Wolfes and Guenther Maass and J{\"u}rgen Alves},
  journal={Gene},
  year={1995},
  volume={157 1-2},
  pages={
          113-8
        }
}
The catalytic center of the restriction endonuclease (ENase) EcoRI is structurally homologous to that of EcoRV, BamHI and PvuII. Each of these ENases contains a short motif of three to four amino acid (aa) residues which are positioned in a similar orientation to the scissile phosphodiester bond. We have mutated these aa (Pro90, Asp91, Glu111 and Lys113) in EcoRI to determine their individual roles in catalysis. The replacement of Asp91 and Lys113, respectively, by conservative mutations (Ala91… CONTINUE READING

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