Rv1626-ARPC4 protein-protein interaction by bacterial two-hybrid.

<p><b>A.</b> Bacterial two-hybrid plate showing Rv1626 and ARPC4 interaction where blue colored colonies show interaction between the bait and target proteins. BacterioMatch two-hybrid <i>E. coli</i> reporter strain was co-transformed with Rv1626-pBTnn+ARPC4-pTRGnn; pBTnn+ARPC4-pTRGnn (negative control); LGF2-pBT+Gal11p-pTRG (positive control); and Rv1626-pBTnn+Gal11p-pTRG (negative control) plasmids. Two individual colonies from each co-transformant were patched on X-gal indicator plate. <b>B.</b> Histogram of liquid β-galactosidase assay for quantitative estimation of interaction strength of Rv1626 and ARPC4 compared to that with positive and negative controls. Co-transformants ARPC4-pTRGnn+pBTnn and Gal11p-pTRG+Rv1626-pBTnn were taken as the negative controls; CFP10-pTRGnn+ESAT6-pBTnn and Gal11p-pTRG+LGF2-pBT were taken as positive controls; and these were used to compare the β-galactosidase activity of co-transformant ARPC4-pTRGnn+Rv1626-pBTnn. Enzyme activity is a direct indication of the strength of interaction of the bait and target proteins and, is expressed in terms of Miller Units. All interactions were found to be statistically significant by Student’s t-test (*, P<0.05).