A transfer vector was constructed in which the neomycin resistance (neo) gene was under the control of a copy of Autographa californica nuclear polyhedrosis virus (AcMNPV) IE1 gene promoter at the p10 locus. After cotransfection of Spodoptera frugiperda (Sf9) cells with the transfer vector and infectious AcMNPV DNA, the recombinant virus-containing neo gene was selected by serial passage of the mixed progenies from cotransfection. This was done at low MOI in the presence of G418 in growth 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.