An in vivo selection system for homing endonuclease activity.
Nucleic Acids Res
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| Abstract | Homing endonucleases are enzymes that catalyze the highly sequence-specific cleavage of DNA. We have developed an in vivo selection in Escherichia coli that links cell survival with homing endonuclease-mediated DNA cleavage activity and sequence specificity. Using this selection, wild-type and mutant variants of three homing endonucleases were characterized without requiring protein purification and in vitro analysis. This selection system may facilitate the study of sequence-specific DNA cleaving enzymes, and selections based on this work may enable the evolution of homing endonucleases with novel activities or specificities. |
| Year of Publication | 2002
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| Journal | Nucleic Acids Res
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| Volume | 30
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| Issue | 7
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| Pages | e29
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| Date Published | 2002 Apr 01
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| ISSN | 1362-4962
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| PubMed ID | 11917035
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| PubMed Central ID | PMC101853
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