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

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.