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 mycobacterial promoter activity. The second vector permits construction of translational fusions of mycobacterial proteins to Gfp in order to study subcellular localization including protein secretion. Using this translational fusion construct, we verify that a Gfp fusion to the putative secreted M. tuberculosis protein ChoD is translocated to the extracellular milieu when cloned and expressed in Mycobacterium smegmatis.
Department of Medicine, Division of Infectious Diseases, University of British Columbia, 2733 Heather St., Vancouver, British Columbia, Canada V5Z 3J5.
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Cowley
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Av-Gay
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D009161:Mycobacterium / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004274:DNA, Recombinant; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins