Evaluating the Sensitivity of Mycobacterium tuberculosis to Biotin Deprivation Using Regulated Gene Expression
Figure 1
Biotin synthesis pathway and construction of Mtb ΔbioA.
(A) Shown are the reactions catalyzed by KAPA synthase (BioF), DAPA synthase (BioA), DTB synthase (BioD) and biotin synthase (BioB), which convert pimeloyl-CoA to biotin, and biotin ligase, which attaches the cofactor to biotin-dependent enzymes. (B) The upper panel displays the genetic organization of the bioA region in wt Mtb; the lower panel displays that of ΔbioA. Gray boxes represent open reading frames of the genes specified above or below, the striped boxes mark the putative bioA promoter, and the black boxes represent hygR. The localization of recognition sites for the restriction endonuclease DraIII are labeled “D”. (C) Southern blot of DraIII-digested genomic DNA from wt Mtb and ΔbioA. The expected sizes for the DraIII fragments hybridizing to the bioF1-derived probe that was used in this blot are 3068 bp for wt and 4021 bp for ΔbioA.