CSIR-CCMB builds fluorescent shuttle vector toolkit to study TB bacterium proteins

Key facts

• Scientists at the CSIR-Centre for Cellular and Molecular Biology (CCMB), led by Director Vinay Kumar Nandicoori, developed a genetic toolkit for studying Mycobacterium tuberculosis, the bacterium that causes TB.
• Research student Asis Kumar created a collection of 40 new shuttle vectors that can carry fluorescent markers such as Green Fluorescent Protein (GFP) and mCherry, making proteins glow under a microscope.
• The toolkit allows scientists to switch genes on, track protein locations and movement inside bacterial cells, and study several proteins simultaneously; four regulatory proteins from the TB bacterium were examined within a single cell.
• The tools were tested in Mycobacterium smegmatis, a harmless laboratory relative of the TB bacterium, where fluorescent tags successfully highlighted proteins involved in cell growth and division.
• The toolkit includes a new purification method that reduces background contamination and improves identification of naturally interacting proteins, and it can be expanded with new genetic switches, protein tags and regulatory elements.
• The study was published in the Journal of Bacteriology.