Giardia lamblia: stable expression of green fluorescent protein mediated by giardiavirus.
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.
Liu
Quan
Q
Department of Veterinary Medicine, JiLin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China.
Zhang
Xichen
X
Li
Jianhua
J
Ying
Jigang
J
Chen
Lifeng
L
Zhao
Yunjun
Y
Wei
Feng
F
Wu
Tao
T
D000818:Animals; D004351:Drug Resistance / Q000235:genetics; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D015967:Gene Expression Regulation, Viral; D016829:Giardia lamblia / Q000235:genetics / Q000378:metabolism* / Q000821:virology*; D019198:Giardiavirus / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D012333:RNA, Messenger; D049452:Green Fluorescent Proteins