Fig. 2.
Analysis of KSHV-lyt replication. (A) Virus reconstitution following BAC transfection into RPE1 cells. GFP expression and focus formation was observed by fluorescence microscopy over time. Images were taken 1 and 8 days posttransfection (dpt). (B) Focus formation 8 days after transferring infected cell supernatant onto fresh RPE1 cells. (C) Comparison of KSHV-lyt BAC and virion DNA. Equal amounts of DNA were digested with PmeI or XhoI and analyzed by gel electrophoresis. Fragments representing the BAC cassette are indicated by arrowheads. Asterisks indicate additional differences. (D) The PmeI and XhoI restriction maps with the expected fragments in the BAC cassette region are depicted. (E) Replication of KSHV-lyt on different cells after low-MOI infection. Titers (focus-forming units) were determined on RPE1 cells using the TCID50 method. DL, detection limit.