An in vivo selection system for homing endonuclease activity
| Autor: | Mathias Gruen, Kathy Chang, Irina Serbanescu, David R. Liu |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2002 |
| Předmět: |
Genetics
Binding Sites Endodeoxyribonucleases biology Mutant DNA Homing endonuclease Cell biology Substrate Specificity Endonuclease chemistry.chemical_compound Plasmid chemistry Mutation biology.protein Escherichia coli Flap endonuclease NAR Methods Online Cell Division Homing (hematopoietic) Plasmids |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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