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NGEC Progress Report Hui Li, Monnat Group

NGEC Progress Report Hui Li, Monnat Group. In vivo and in vitro cleavage specificities. Construction of degenerate site library for I-CreI and I-MsoI. I-CreI library: ~ 10 7 unique sites; 6.8 ± 2.5 mutations/per site I-MsoI library: ~ 5 × 10 6 unique sites; 8.2 ± 2.1 mutations/per site.

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NGEC Progress Report Hui Li, Monnat Group

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  1. NGEC Progress ReportHui Li, Monnat Group • In vivo and in vitro cleavage specificities • Construction of degenerate site library for I-CreI and I-MsoI I-CreI library: ~ 107 unique sites; 6.8 ± 2.5 mutations/per site I-MsoI library: ~ 5 ×106 unique sites; 8.2 ± 2.1 mutations/per site • Construction of single bp mismatch matrix for I-MsoI In vivo cleavage specificity based on recombination In vitro cleavage specificity • Generation of Mso mutants with higher activity • rational design • bacterial selection

  2. In vivo cleavage specificity of mMsoI

  3. In vitro cleavage specificity of Mso-I Each target site 7.5 nm

  4. In vitro cleavage specificity of mMsoI Each target site 7.5 nm

  5. Rational design of I-MsoI Mutants D81 D146, E147 E147 D146 J. M. B. (2003) 329, 253-269

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