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 expressio n of heterologous proteins in either dipteran or lepidopteran insect c ell lines. Constitutive expression in a broad range of host cells is m ediated by the Orgyia pseudotsugata multicapsid nucleopolyhedrosis vir us (OpMNPV) immediate-early 2 (ie2) promoter. Alternatively, if induci ble expression is required, for example to express cytotoxic proteins, a vector has been constructed that uses the Drosophila metallothionei n (Mtn) promoter for metal-inducible protein expression in dipteran ce ll lines. A chimeric synthetic bacterial-OpMNPV ie promoter-Zeocin(R) resistance gene cassette has been included to facilitate cloning in E. coli as well as the generation of stably transformed insect cell line s. The utility of the system is demonstrated by the constitutive and i nducible expression of the highly processed glycosylphosphatidylinosit ol-anchored glycoprotein, human melanotransferrin, in transformed inse ct cell lines. (C) 1998 Elsevier Science B.V.