Substrate and target sequence length influence RecTE(Psy) recombineering efficiency in Pseudomonas syringae.
PLoS One
; 7(11): e50617, 2012.
Article
em En
| MEDLINE
| ID: mdl-23226333
ABSTRACT
We are developing a new recombineering system to assist experimental manipulation of the Pseudomonas syringae genome. P. syringae is a globally dispersed plant pathogen and an important model species used to study the molecular biology of bacteria-plant interactions. We previously identified orthologs of the lambda Red bet/exo and Rac recET genes in P. syringae and confirmed that they function in recombineering using ssDNA and dsDNA substrates. Here we investigate the properties of dsDNA substrates more closely to determine how they influence recombineering efficiency. We find that the length of flanking homologies and length of the sequences being inserted or deleted have a large effect on RecTE(Psy) mediated recombination efficiency. These results provide information about the design elements that should be considered when using recombineering.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Recombinação Genética
/
Engenharia Genética
/
Pseudomonas syringae
/
Genes Bacterianos
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article