AN EFFICIENT METHOD TO IDENTIFY POSITIVE RECOMBINANT BACULOVIRUS BY USING NEO AS SELECTION MARKER
以neo作选择标记富集和筛选阳性重组杆状病毒

A cassette of neomycin resistance gene under the AcNPV IE1 gene promotorwas inserted into a p10-based transfer vector pAcEP106. Recombinant plasmidpAcPIneo was selected and cotransfected Sf cells with wild type AcNPV DNA.Expression of the neo gene under IE1 promotor enable recombinant virus multiply in theSf cells treated with antibiotic G418 which inhibit the wild-type virus growth. Passagethe cotransfection suspend at low MOI, the proportion of neo-containing virusesincrease. After three passages, the recombinants accounted for over 90%. This work laya foundation for using baculovirus early gene promoter to drive foreign gene expressionand also established a simplified procedure for selection of positive recombinantbaculoviruses.