A novel expression system based on host-range expansion of baculovirus

A host range expanded recombinant Autographa californica multiple-nucleocap sid nucleopolyhedrosis virus AcM-NPV/r2 was obtained by cotransfection of t he bacmid DNA from Escherichia coli DH10Bac along with a plasmid pBmH-M con taining HindIII M fragment of Bombyx mori nuclear polyhedrosis Virus (BmNPV ) genomic DNA. A recombinant transposon vector carrying a mutant green fluo rescent protein gene (GFP) and a polyhedrin gene was constructed. Transposi tion was carried out in both E. coli DH10Bac and E. coli DH10BmH, which con tains AcMNPV/r2 and a helper plasmid. Recombinant DNAs were transfected int o Sf-9 cells to generate recombinant virus AcMNPV/r3 and AcMNPV/r4 respecti vely. Viral stock of AcMNPV/r4 was then infected into Bombyx mori cells (Bm N) and Bombyx,mori larvae (silkworm). Analysis shows that GFP was highly ex pressed in Bombyx mori larvae. This expression system, is practicable there fore for mass production of foreign gene products. (C) 1998 Elsevier Scienc e B.V. All rights reserved.