Facile construction of unmarked deletion mutants in Burkholderia pseudomallei using sacB counter-selection in sucrose-resistant and sucrose-sensitive isolates

J Microbiol Methods. 2009 Mar;76(3):320-3. doi: 10.1016/j.mimet.2008.12.007. Epub 2008 Dec 29.


Burkholderia pseudomallei is the causative agent of melioidosis, a potentially fatal disease endemic or emerging world-wide. Here we report unmarked allele-replacement mutagenesis using efficient sacB counter-selection. Despite being genotypically sacB(+), most commonly used B. pseudomallei strains are sucrose-resistant and efficient sacB counter-selection is demonstrated in both resistant and sensitive strains.

MeSH terms

  • Alleles
  • Burkholderia pseudomallei / genetics*
  • Burkholderia pseudomallei / metabolism
  • DNA, Bacterial / genetics
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial
  • Mutagenesis, Site-Directed / methods*
  • Sequence Deletion
  • Sucrose / metabolism*


  • DNA, Bacterial
  • Sucrose