Cytosine deaminase as a negative selection marker for gene disruption and replacement in the genus Streptomyces and other actinobacteria

Appl Environ Microbiol. 2009 Feb;75(4):1211-4. doi: 10.1128/AEM.02139-08. Epub 2008 Dec 19.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actinobacteria / genetics*
  • Anti-Bacterial Agents / pharmacology
  • Cytosine Deaminase / genetics
  • Cytosine Deaminase / metabolism*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Flucytosine / metabolism
  • Flucytosine / pharmacology
  • Fluorouracil / metabolism
  • Fluorouracil / pharmacology
  • Genetic Vectors
  • Microbial Sensitivity Tests
  • Mutagenesis, Insertional / methods*
  • Selection, Genetic*

Substances

  • Anti-Bacterial Agents
  • Escherichia coli Proteins
  • Flucytosine
  • Cytosine Deaminase
  • codA protein, E coli
  • Fluorouracil