Open AccessCytosine Deaminase as a Negative Selection Marker for Gene Disruption and Replacement in the Genus Streptomyces and Other Actinobacteria
Author(s) -
Marie-Pierre Dubeau,
Mariana Gabriela Ghinet,
Pierre-Étienne Jacques,
Nancy Clermont,
Carole Beaulieu,
Ryszard Brzeziński
Publication year - 2009
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.02139-08
Subject(s) - actinobacteria , biology , streptomyces , cytosine deaminase , gene , genetics , cytosine , escherichia coli , selection (genetic algorithm) , cytidine deaminase , bacteria , 16s ribosomal rna , genetic enhancement , computer science , artificial intelligence
We developed a novel negative selection system for actinobacteria based on cytosine deaminase (CodA). We constructed vectors that include a synthetic gene encoding the CodA protein fromEscherichia coli optimized for expression inStreptomyces species. Gene disruption and the introduction of an unmarked in-frame deletion were successfully achieved with these vectors.