An in vivo selection system for homing endonuclease activity
Open Access
- 1 April 2002
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 30 (7) , 29e-29
- https://doi.org/10.1093/nar/30.7.e29
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.Keywords
This publication has 27 references indexed in Scilit:
- Chemical Probing Shows That the Intron-encoded Endonuclease I-SceI Distorts DNA through Binding in Monomeric Form to Its Homing SiteJournal of Biological Chemistry, 2001
- Homing endonucleases: keeping the house in orderNucleic Acids Research, 1997
- Characterization of an ‘orthogonal’ suppressor tRNA derived from E. coli tRNA2GlnChemistry & Biology, 1997
- [17] Fluorescence assays for DNA cleavagePublished by Elsevier ,1997
- In vivo system for the detection of low level activity barnase mutantsProtein Engineering, Design and Selection, 1995
- A Fast and Accurate Enzyme-Linked Immunosorbent Assay for the Determination of the DNA Cleavage Activity of Restriction EndonucleasesAnalytical Biochemistry, 1993
- Continuous spectrophotometric assay for restriction endonucleases using synthetic oligodeoxynucleotides and based on the hyperchromic effectAnalytical Biochemistry, 1992
- Barnase and barstar: two small proteins to fold and fit togetherTrends in Biochemical Sciences, 1989
- Barnase and barstarJournal of Molecular Biology, 1988
- Effects of functional group changes in the EcoRI recognition site on the cleavage reaction catalyzed by the endonucleaseBiochemistry, 1987