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[Giardia canis virus transfection vector-mediated expression of green fluorescent protein in the parasite]. | LitMetric
Objective : To construct Giardia canis virus (GCV) transfection vector.Methods : According to transcriptional start site, replication origin and packaging site of GCV genome (DQ238861), a system was developed for the expression of a foreign gene in this organism by flanking the green fluorescent protein (GFP) gene with the fragments of GCV positive-strand RNA. The transcript of the construct was synthesized in vitro with T7 RNA polymerase and used to transfect GCV-infected trophozoites by electroporation.Results : The recombinant plasmid pGCV634/GFP/GCV2174 was constructed. The expression of green fluorescent protein mediated by GCV transfection vector in Giardia canis peaked at 1 d after electroporation (A490=1.8), and slowly decreased until 14 d post-transfection.Conclusion : The engineered GCV vector can be successfully used to introduce and efficiently express a heterologous gene in the eukaryotic microorganism.
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