Protocol for confocal microscopy-based localization of chorismate mutase protein of Mycobacterium tuberculosis in trans-Golgi compartment.

Here, we present a confocal microscopy protocol to determine the localization of Mycobacterium tuberculosis chorismate mutase (MtbCM) in trans-Golgi compartment. We co-transfect HEK293 cells with either pEGFP-C1-MtbCM and mCherry-TGOLN2 (trans-Golgi marker) or pEGFP-C1 and mCherry-TGOLN2 as a control, using Lipofectamine 2000. We describe steps for constructing pEGFP-C1-MtbCM and performing co-transfection and image acquisition using confocal microscopy. This protocol provides a representative procedure to determine the subcellular localization of the mycobacterial protein to the trans-Golgi compartment. For complete details on the use and execution of this protocol, please refer to Pal et al.1.