A series of broad host range shuttle vectors for constitutive and inducible expression of heterologous proteins in insect cell lines.
A series of shuttle vectors have been constructed that allow expression of heterologous proteins in either dipteran or lepidopteran insect cell lines. Constitutive expression in a broad range of host cells is mediated by the Orgyia pseudotsugata multicapsid nucleopolyhedrosis virus (OpMNPV) immediate-early 2 (ie2) promoter. Alternatively, if inducible expression is required, for example to express cytotoxic proteins, a vector has been constructed that uses the Drosophila metallothionein (Mtn) promoter for metal-inducible protein expression in dipteran cell lines. A chimeric synthetic bacterial-OpMNPV ie promoter-Zeocin resistance gene cassette has been included to facilitate cloning in E. coli as well as the generation of stably transformed insect cell lines. The utility of the system is demonstrated by the constitutive and inducible expression of the highly processed glycosylphosphatidylinositol-anchored glycoprotein, human melanotransferrin, in transformed insect cell lines.
Hegedus
D D
DD
Department of Zoology, University of British Columbia, Vancouver, Canada.
Pfeifer
T A
TA
Hendry
J
J
Theilmann
D A
DA
Grigliatti
T A
TA
D000818:Animals; D000951:Antigens, Neoplasm; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D004330:Drosophila; D004926:Escherichia coli; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D007313:Insecta / Q000166:cytology; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins / Q000235:genetics; D017924:Nucleopolyhedroviruses; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics; D018411:Spodoptera
D000951:Antigens, Neoplasm; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D011994:Recombinant Proteins