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Development of a Negative Selectable Marker for Entamoeba histolytica
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December 12, 2010
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We report development of a negative selection system in E. histolytica based upon transgenic expression of a chimeric protein (FCU1) and selection with the prodrug 5-fluorocytosine. The FCU1 protein is a fusion of yeast cytosine deaminase and uracil phosphoribosyltransferase. Expression of FCU1 resulted in increased E. histolytica sensitivity towards 5-fluorocytosine.
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Negative Selectable MarkerEntamoeba HistolyticaAmebiasisGene ExpressionTransfectionPlasmidsReverse GeneticsHomologous RecombinationEpisomal Plasmid DNABackground Of Exogenous DNAGenomic Integration EventNegative Selection SystemYeast Cytosine Deaminase And Uracil Phosphoribosyl Transferase Chimera (FCU1)Prodrug 5-fluorocytosine (5-FC)5-fluorouracil5-fluorouridine-5′-monophosphate
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Abhyankar, M. M., Haviland, S. M., Gilchrist, C. A., Petri, Jr., W. A. Development of a Negative Selectable Marker for Entamoeba histolytica. J. Vis. Exp. (46), e2410, doi:10.3791/2410 (2010).
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