In vivo determination of the gap2 gene promoter activity in Giardia lamblia

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MeSH: Animals ; Blotting, Southern/methods ; GTPase-Activating Proteins/genetics* ; Gene Expression/genetics ; Gene Expression/physiology* ; Gene Order ; Genes, Protozoan/genetics ; Genes, Protozoan/physiology* ; Genes, Reporter/genetics ; Genetic Engineering/methods ; Giardia lamblia/genetics* ; Life Cycle Stages/physiology ; Luciferases/genetics ; Luciferases/metabolism ; Plasmids ; Promoter Regions, Genetic/physiology* ; Recombinant Fusion Proteins/analysis ; Recombinant Fusion Proteins/biosynthesis ; Time Factors ; Transfection/methods

Keywords: Giardia lamblia ; gap2 gene expression ; encystation ; luciferase ; transfection

Abstract: A shuttle vector for Escherichia coli and Giardia lamblia was modified to produce a reporter plasmid, which monitors the expression of prescribed gene in G. lamblia by measuring its luciferase activity. Promoter regions of the gap2 gene, one of the genes induced during encystation, were cloned into this plasmid, and the resultant constructs were then transfected into trophozoites of G. lamblia. Transgenic trophozoites containing one of the 3 gap2-luc reporters were induced to encystation, and characterized with respect to gap2 gene expression by measuring their luciferase activities. Giardia containing a gap2-luc fusion of 112-bp upstream region showed full induction of luciferase activity during encystation.