Purification and In Vitro Characterization of Zinc Finger Recombinases.
Methods Mol Biol
; 1642: 229-245, 2017.
Article
em En
| MEDLINE
| ID: mdl-28815504
Zinc finger recombinases (ZFRs) are designer site-specific recombinases that have been adapted for a variety of genome editing purposes. Due to their modular nature, ZFRs can be customized for targeted sequence recognition and recombination. There has been substantial research on the in vivo properties and applications of ZFRs; however, in order to fully understand and customize them, it will be necessary to study their properties in vitro. Experiments in vitro can allow us to optimize catalytic activities, improve target specificity, measure and minimize off-target activity, and characterize key steps in the recombination pathway that might be modified to improve performance. Here, we present a straightforward set of protocols for the expression and purification of ZFRs, an assay system for catalytic proficiency in vitro and bandshift assays for detection of sequence-specific DNA interactions.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
DNA
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Dedos de Zinco
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Clonagem Molecular
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Ensaio de Desvio de Mobilidade Eletroforética
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DNA Nucleotidiltransferases
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Escherichia coli
Limite:
Humans
Idioma:
En
Revista:
Methods Mol Biol
Ano de publicação:
2017
Tipo de documento:
Article