Expression of the green fluorescent protein (GFP) in mycobacterium avium as a tool to study the interaction between Mycobacteria and host cells.

TitleExpression of the green fluorescent protein (GFP) in mycobacterium avium as a tool to study the interaction between Mycobacteria and host cells.
Publication TypeJournal Article
Year of Publication1997
AuthorsParker AE, Bermudez LE
JournalMicrobial pathogenesis
Volume22
Issue4
Pagination193-8
Date Published1997 Apr
ISSN0882-4010
KeywordsCells, Cultured, Cloning, Molecular, Gene Expression, Genetic Vectors, Green Fluorescent Proteins, HT29 Cells, Humans, Luminescent Proteins, Macrophages, Mycobacterium avium, Plasmids, Recombinant Fusion Proteins, Transformation, Genetic, Virulence
Abstract

Mycobacterium avium is an intracellular pathogen able to invade and survive within macrophages and mucosal epithelial cells. The study of the interaction between M. avium and host cells is important to establish the mechanisms of pathogenesis of the infection. One of the limitations of microscopic study of intracellular M. avium is the difficulty of identifying isolated bacilli within cells. As a general strategy to visualize and analyse the influence of M. avium on intracellular trafficking, we cloned the gene encoding the green fluorescent protein (GFP) in M. avium. A vector was constructed by cloning the cDNA for the GFP of the bioluminescent jellyfish Aequorea victoria into an Escherichia coli/Mycobacteria shuttle vector (pMV261). The recombinant plasmid (pWES-4) was transformed into both E. coli and M. avium strain 104 (serovar 1). The transformants were screened for strong expression of the GFP. Transformed M. avium were clearly visible inside human macrophages and epithelial cells using fluorescence microscopy. These transformed M. avium should provide a useful tool for further study of intracellular behaviour.

DOI10.1006/mpat.1996.0106
Alternate JournalMicrob. Pathog.