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. 2011 Nov 8;6(11):e27503. doi: 10.1371/journal.pone.0027503

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Tharad et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) X-Gal indicator plates (with and without arabinose) showing triple co-transformants: ESAT6pBTnn + SL3pTRGnn + pMTSA; ESAT6pBTnn + HCL1pTRGnn + pMTSA; ESAT6pBTnn + SL3pTRGnn + CFP10pMTSA; and ESAT6pBTnn + HCL1pTRGnn + CFP10pMTSA. Two different colonies of each co-transformant were patched. (B) Fold-difference of lacZ gene transcript in the triple co-transformants relative to the ESAT6pBTnn + HCL1pTRGnn + pMTSA negative control strain. The lacZ gene transcription is reduced approx. four-fold as compared to negative control when ESAT6 and HCL1 interact in presence of CFP10 in the bacterial three-hybrid system (*, P<0.05), whereas no significant difference is observed in the ESAT6 : SL3 strain expressing CFP10 (**, P>0.05). The RT-PCR experiment was performed in triplicate using 3 separate colonies.