HIGHLY EFFICIENT PRODUCTION OF GFP AND ITS DERIVATIVES IN INSECT CELLS FOR VISUAL IN-VITRO APPLICATIONS

We have generated recombinant baculoviruses for expression of the gree n fluorescent protein (GFP), a bright GFP mutant (S65T), and a GFP-str eptavidin fusion protein in Sf9 and High Five insect cell lines, At 3- 4 days post infection, about 30% of the total protein contents was rep resented by the recombinant protein products, giving the infected inse ct cells a bright green color which was clearly visible by eye in dayl ight, The isolated GFP-streptavidin fusion protein, which possessed fl uorescence properties identical to those of the native GFP, was capabl e of binding biotin as shown by using biotinylated beads as well as bi otinylated antibody complexes decorating surface expressed GluR-6 glut amate receptor in live and fixed insect cells. The exceptionally high expression levels of GFP and GFP (S65T) and the GFP-streptavidin fusio n protein in recombinant baculovirus infected insects should facilitat e production of GFP derivatives for in vitro applications.