Evaluating the Sensitivity of Mycobacterium tuberculosis to Biotin Deprivation Using Regulated Gene Expression
Figure 2
In vitro characterization of Mtb ΔbioA.
(A) Growth of ΔbioA in Sauton's medium (left) or Sauton's medium supplemented with KAPA, DTB or biotin. Data are from individual cultures and representative of at least two independent experiments. (B)Growth of ΔbioA in Sauton's medium with varying concentrations of biotin. Data are from individual cultures and representative of at least two independent experiments. (C) Immunoblots performed with both an anti-biotin antiserum and rabbit serum recognizing Mtb's dihydrolipoamide acyltransferase (DlaT). The lower panel shows the signal from the anti-biotin antibody; the upper panel shows the DlaT signal, which served as the loading control. Protein extracts were prepared after the indicated days of cultivation in biotin-free Sauton's medium. Data are from individual cultures and representative of at least two independent experiments. (D) Survival of wt and ΔbioA in Sauton's media with and without biotin. Data are averages from triplicate cultures and representative of at least two independent experiments. Error bars indicate standard deviations.