title: Cytosine deaminase as a negative selection marker for gene disruption and replacement in the genus Streptomyces and other actinobacteria.
abstract: 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 from Escherichia coli optimized for expression in Streptomyces species. Gene disruption and the introduction of an unmarked in-frame deletion were successfully achieved with these vectors.
authors.affiliation: Centre d'Etude et de Valorisation de la Diversité Microbienne, Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, QC J1K 2R1, Canada.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Dubeau
authors.forename: Mariana Gabriela
authors.initials: MG
authors.lastname: Ghinet
authors.forename: Pierre-Etienne
authors.initials: PE
authors.lastname: Jacques
authors.forename: Nancy
authors.initials: N
authors.lastname: Clermont
authors.forename: Carole
authors.initials: C
authors.lastname: Beaulieu
authors.forename: Ryszard
authors.initials: R
authors.lastname: Brzezinski
mesh_terms: D039903:Actinobacteria / Q000235:genetics*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D043525:Cytosine Deaminase / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005437:Flucytosine / Q000378:metabolism / Q000494:pharmacology; D005472:Fluorouracil / Q000378:metabolism / Q000494:pharmacology; D005822:Genetic Vectors; D008826:Microbial Sensitivity Tests; D016254:Mutagenesis, Insertional / Q000379:methods*; D012641:Selection, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D005437:Flucytosine; D043525:Cytosine Deaminase; C468827:codA protein, E coli; D005472:Fluorouracil