A BACULOVIRUS VECTOR DERIVED FROM IMMEDIATELY EARLY GENE PROMOTER OF AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS

A transfer vector was constructed in which the neomycin resistance (ne o) gene was under the control of a copy of Autographa californica nucl ear polyhedrosis virus (AcMNPV) IE1 gene promoter at the p10 locus. Af ter cotransfection of Spodoptera frugiperda (Sf9) cells with the trans fer vector and infectious AcMNPV DNA, the recombinant virus-containing neo gene was selected by serial passage of the mixed progenies from c otransfection. This was done at low MOI in the presence of G418 in gro wth medium and was followed by limited dilution. RNA dot hybridization showed that the neo gene was transcribed from immediately early phase to very late phase, in infected Sf9 cells. These results demonstrate that a new baculovirus vector system had been constructed in infected cells. Furthermore, a new method for selection of positive recombinant baculovirus had been developed.