In vivo determination of the gap2 gene promoter activity in Giardia lamblia.
A shuttle vector for Escherichia coli and Giardia lamblia was modified to produce a reporter plasmid, which monitors the expression of prescribed gene in G. lamblia by measuring its luciferase activity. Promoter regions of the gap2 gene, one of the genes induced during encystation, were cloned into this plasmid, and the resultant constructs were then transfected into trophozoites of G. lamblia. Transgenic trophozoites containing one of the 3 gap2-luc reporters were induced to encystation, and characterized with respect to gap2 gene expression by measuring their luciferase activities. Giardia containing a gap2-luc fusion of 112-bp upstream region showed full induction of luciferase activity during encystation.
Yang
Hye-Won
HW
Department of Parasitology and Institute of Tropical Medicine, Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Korea.
Kim
Juri
J
Yong
Tai-Soon
TS
Park
Soon-Jung
SJ
D000818:Animals; D015139:Blotting, Southern / Q000379:methods; D020690:GTPase-Activating Proteins / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics / Q000502:physiology*; D023061:Gene Order; D017125:Genes, Protozoan / Q000235:genetics / Q000502:physiology*; D017930:Genes, Reporter / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D016829:Giardia lamblia / Q000235:genetics*; D008018:Life Cycle Stages / Q000502:physiology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis; D013997:Time Factors; D014162:Transfection / Q000379:methods
D020690:GTPase-Activating Proteins; D011993:Recombinant Fusion Proteins; D008156:Luciferases