Expression of the green fluorescent protein (GFP) inMycobacterium aviumas a tool to study the interaction betweenMycobacteriaand host cells

• Published in: Microbial pathogenesis Vol. 22; no. 4; pp. 193 - 198
• Main Authors: Parker, A.E., Bermudez, L.E.
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01.04.1997
• Subjects: green fluorescent protein (GFP); intracellular pathogen; microbial pathogenesis; Mycobacterium avium; Mycobacterium avium; microbial pathogenesis; green fluorescent protein (GFP); intracellular pathogen
• ISSN: 0882-4010; 1096-1208
• DOI: 10.1006/mpat.1996.0106

Mycobacterium aviumis an intracellular pathogen able to invade and survive within macrophages and mucosal epithelial cells. The study of the interaction betweenM. aviumand host cells is important to establish the mechanisms of pathogenesis of the infection. One of the limitations of microscopic study of intracellularM. aviumis the difficulty of identifying isolated bacilli within cells. As a general strategy to visualize and analyse the influence ofM. aviumon intracellular trafficking, we cloned the gene encoding the green fluorescent protein (GFP) inM. avium. A vector was constructed by cloning the cDNA for the GFP of the bioluminescent jellyfishAequorea victoriainto anEscherichia coli/Mycobacteriashuttle vector (pMV261). The recombinant plasmid (pWES-4) was transformed into bothE. coliandM. aviumstrain 104 (serovar 1). The transformants were screened for strong expression of the GFP. TransformedM. aviumwere clearly visible inside human macrophages and epithelial cells using fluorescence microscopy. These transformedM. aviumshould provide a useful tool for further study of intracellular behaviour.