[Giardia canis virus transfection vector-mediated expression of green fluorescent protein in the parasite].
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.


CONCLUSIONS
The engineered GCV vector can be successfully used to introduce and efficiently express a heterologous gene in the eukaryotic microorganism.
Chen
Li-Feng
LF
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China.
Li
Jian-Hua
JH
Liu
Quan
Q
Zhao
Yue-Ping
YP
Cao
Li-Li
LL
Zhang
Xi-Chen
XC
D000818:Animals; D002478:Cells, Cultured; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics; D005872:Giardia / Q000166:cytology / Q000235:genetics* / Q000821:virology; D019198:Giardiavirus / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins