Cytosine deaminase as a negative selection marker for gene disruption and replacement in the genus Streptomyces and other actinobacteria.
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.
Dubeau
Marie-Pierre
MP
Centre d'Etude et de Valorisation de la Diversité Microbienne, Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, QC J1K 2R1, Canada.
Ghinet
Mariana Gabriela
MG
Jacques
Pierre-Etienne
PE
Clermont
Nancy
N
Beaulieu
Carole
C
Brzezinski
Ryszard
R
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
D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D005437:Flucytosine; D043525:Cytosine Deaminase; C468827:codA protein, E coli; D005472:Fluorouracil