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Figure 1 | BMC Infectious Diseases

Figure 1

From: A peptide fragment from the human COX3 protein disrupts association of Mycobacterium tuberculosisvirulence proteins ESAT-6 and CFP10, inhibits mycobacterial growth and mounts protective immune response

Figure 1

HCL2 binds with ESAT-6 and disrupts the ESAT-6:CFP10 heterodimeric complex. (A) Agar plate showing co-transformants: a – ‘white’ negative control pBTnn + ESAT-6pTRGnn; b – interacting clone ESAT-6pBTnn + HCL2pTRGnn; c – ‘blue’ positive control CFP10pTRGnn + ESAT-6pBTnn. (B) Interactions between ESAT-6pBTnn + HCL2pTRGnn and ESAT-6pBTnn + CFP10pTRGnn were found comparable by Liquid β-Galactosidase assay. Experiments were repeated thrice and similar observations were made. All readings were found statistically significant by applying Student’s T-test. (*P < 0.05) (C) HCL2-His6X interacts with ESAT-6-FLAG in vitro. Experiment was repeated thrice and each time positive interaction was observed between HCL2-His6X and ESAT-6-FLAG. (D) Disruption of ESAT-6:CFP10 interaction by HCL2 using the three-hybrid system. Three separate colonies from each triple co-transformants plate were picked and patched. a – (ESAT-6pBTnn-CFP10pTRGnn) + HCL2pMTSA; b – (ESAT-6pBTnn-CFP10pTRGnn) + pMTSA. –ARA represents plate without Arabinose induction while + ARA represents plate with Arabinose induction. Experiments were repeated thrice and similar observations were made. (E) Bacterial Three-Hybrid results were also corroborated by Arabinose Gradient Liquid β-Galactosidase Assay. Experiment was repeated thrice in triplicates. All readings were found statistically significant by applying Student’s T-test (*P < 0.05).

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BMC Infectious Diseases

ISSN: 1471-2334

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