Stable expression of green fluorescent protein mediated by GCV in Giardia canis

Giardia canis can be infected with a double-stranded RNA virus, that is giardiavirus (G. canis virus, GCV). In this study, green fluorescent protein (GFP) was stably expressed in G. canis mediated by GCV. The plasmid pNEO/GDH/MCS/GFP, containing the neomycin phosphotransferase (NEO) encoding region flanked by the 636 nt of 5'-terminus and the 2174 nt of 3'-terminus from GCV positive strand RNA, was constructed by inserting GFP gene into downstream from the NEO gene and glutamate dehydrogenase (GDH) 5'-terminus uncoding regions on a single plasmid, and its in vitro transcript was introduced into GCV-infected G. canis by electroporation. The transfectants expressed GFP persistently under G418 selection. This stable transfection system should provide a valuable tool for genetic study of G. canis.