Representation of ESAT6 : HCL1 protein-protein disruption by CFP10 in bacterial three-hybrid system.
(A) X-Gal indicator plate with and without arabinose patched with ESAT6pBTnn + HCL1pTRGnn + pMTSA; and ESAT6pBTnn + HCL1pTRGnn + CFP10pMTSA. Blue colony turns white when CFP10 is allowed to express in the presence of 1% arabinose. (B) Time course liquid β-galactosidase assay: β-galactosidase activity of above triple co-transformants: (▪) ESAT6pBTnn + HCL1pTRGnn + pMTSA; and (•) ESAT6pBTnn + HCL1pTRGnn + CFP10pMTSA is plotted against time-points of bacterial culture growth with 0 time-point being the point of arabinose induction. Standard deviation of the activities obtained in three separate assays is shown by error bars. (P<0.05 at all time-points beyond 60 minutes) (C) Arabinose gradient liquid β-galactosidase assay: Relative β-galactosidase activity of triple co-transformants (•) ESAT6pBTnn + HCL1pTRGnn + CFP10pMTSA; and (▪) ESAT6pBTnn + HCL1pTRGnn + pMTSA is plotted against arabinose concentration. The graph is the average of three separate assays and standard deviation is represented as error bars. (P<0.05 at all arabinose concentrations beyond 0.01%).