Massively parallel determination and modeling of endonuclease substrate specificity.
Nucleic Acids Res
; 42(22): 13839-52, 2014 Dec 16.
Article
em En
| MEDLINE
| ID: mdl-25389263
ABSTRACT
We describe the identification and characterization of novel homing endonucleases using genome database mining to identify putative target sites, followed by high throughput activity screening in a bacterial selection system. We characterized the substrate specificity and kinetics of these endonucleases by monitoring DNA cleavage events with deep sequencing. The endonuclease specificities revealed by these experiments can be partially recapitulated using 3D structure-based computational models. Analysis of these models together with genome sequence data provide insights into how alternative endonuclease specificities were generated during natural evolution.
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1
Base de dados:
MEDLINE
Assunto principal:
Endodesoxirribonucleases
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article