Journal: Scientific Reports

Article Title: Cyclic di-GMP regulates Mycobacterium tuberculosis resistance to ethionamide

doi: 10.1038/s41598-017-06289-7

Figure Lengend Snippet: Characterization of the interaction between Ethr and c-di-GMP. ( a ). c-di-GMP binds protein Ethr on the protein microarray. Cy3 signal was measured at 532 nm by a fluorescence microarray scanner. +: in the presence of biotin-c-di-GMP; −: in the presence of biotin alone. ( b ). UV cross-linking assay for validation the interaction between Ethr and c-di-GMP. Purified GST tag Ethr was co-incubated with 1, 5, 25 μM biotinylated c-di-GMP or 25 μM biotinylated cGMP or 25 μM biotinylated c-di-GMP (Bio-c-di-GMP) with 1000 μM c-di-GMP competition. The input was visualized by a GST antibody. The relative signal to control is presented below. ( c ). Measure the binding kinetics of c-di-GMP and Ethr by Bio-Layer interference (BLI). Bio-c-di-GMP was immobilized on streptavidin coated biosenser and incubated with serially diluted Ethr. The measurement and calculation were carried out according to the manufacturer’s instruction.

Article Snippet: Biotin-c-di-GMP, Biotin-c-di-AMP and Biotin-c-GMP were purchased from BioLog Life Science Institute, Bremen, Germany.

Techniques: Microarray, Fluorescence, Purification, Incubation, Binding Assay