EXPRESSION OF GREEN FLUORESCENT PROTEIN WITH BACULOVIRUS VECTOR IN INSECT CELLS
绿色荧光蛋白基因在昆虫细胞中的克隆与表达

The green fluorescent protein (GFP) gene was subcloned into the transfer vector pVLneo downstream of the polyhedrin gene (ocu) promoter. Insect cells were cotransfected with recombinant plasmid and Autographa califomica Nuclear Polyhedrosis Virus (AcNPV) DNA. In the presence of G418, the recombinant virus containing GFP gene was purified. The GFP expressed in insect cells with a Mw of SOkDa is observable by strong green light under a fluorescent microscope. Excitation and emission spectra of the GFP were 395nm and 509nm respectively. Integration of GFP gene on AcNPV genome was identified directly by Southern blot which gave strong hybridization signal between GFP cDNA probe and 1kb EcoRI fragment of recombinant virus.