Monitoring promoter activity and protein localization in Mycobacterium spp. using green fluorescent protein

Two green fluorescent protein (Gfp) fusion Vectors were constructed for use in Mycobacterium spp. The first plasmid facilitates quantification of myco bacterial promoter activity. The second vector permits construction of tran slational fusions of mycobacterial proteins to Gfp in order to study subcel lular localization including protein secretion. Using this translational fu sion construct, we verify that a Gfp fusion to the putative secreted M. tub erculosis protein ChoD is translocated to the extracellular milieu when clo ned and expressed in Mycobacterium smegniatis. (C) 2001 Elsevier Science B. V. All rights reserved.