FIG. 3.
Analysis of viral replication by a lef-11-null BACmid. (A) A transfection-infection assay was used to examine lef-11-null BACmids for viral replication in Sf9 cells. BACmid DNAs from the indicated constructs were used to transfect Sf9 cells, and cells were incubated for 5 days. Supernatants from transfected cells were transferred to a second group of Sf9 cells, which were subsequently incubated for 3 days and then stained for GUS expression from the p6.9 late promoter-GUS reporter. The results of GUS staining are shown in the three lower panels. (B) Virus growth curves were determined from either transfected cells or infected cells. For transfections, Sf9 cells were transfected in triplicate with either vAclef11KO or vAc64−/+GUS, and then supernatants were removed at the indicated times posttransfection, and the TCID50 in Sf9 cells was determined. For infections, infectious budded virions were prepared from wild-type AcMNPV or BACmids vAclef11KO-REP or vAc64−/+GUS. Infections were performed at an MOI of 5 in triplicate, and supernatants were collected and assayed for production of infectious virus by TCID50.