Giardia lamblia: stable expression of green fluorescent protein mediated by giardiavirus

• DOI: 10.1016/j.exppara.2004.12.006
• Publication Date: 2005-01-25T18:14:04Z
• Authors: Quan Liu; Xichen Zhang; Jianhua Li; Jigang Ying; Lifeng Chen; YunJun Zhao; Feng Wei; Tao Wu
• Keywords: Gene Expression Regulation, Viral; 0301 basic medicine; 0303 health sciences; Reverse Transcriptase Polymerase Chain Reaction; Green Fluorescent Proteins; Drug Resistance; Enzyme-Linked Immunosorbent Assay; Transfection; 3. Good health; Giardiavirus; 03 medical and health sciences; Electroporation; Microscopy, Fluorescence; Animals; RNA, Messenger; Giardia lamblia; Plasmids

Giardia lamblia, an early diverging eukaryote that infects several species including humans and a major agent of water-borne diarrhea throughout the world, can be infected with a double-stranded RNA virus, giardiavirus (GLV). A chimeric GLV cDNA and green fluorescent protein (GFP) according to the cis-acting signals of the GLV genome required for expression of foreign gene was constructed and its in vitro transcript was electroporated into GLV-infected G. lamblia trophozoites, GFP was expressed transiently. pGDH5/NEO/GLV was constructed by combining the neomycin resistance cassette in which the neomycin phosphotransferase gene was flanked by Giardia glutamate dehydrogenase (GDH) uncoding regions and the transcription cassette in which the chimera of GLV cDNA and GFP was located downstream from GDH gene promoter on a single plasmid. This plasmid was electroporated into G. lamblia and the transfectants persistently expressed GFP under G418 selection. This stable transfection system should provide a valuable tool for genetic study of G. lamblia.