Epstein-Barr Virus Recombinants: Use of Positive Selection Markers to Rescue Mutants in EBV Negative B Lymphoma Cells

Abstract

Epstein-Barr virus (EBV) genetic analyses have been restricted because the in vitro host range is limited to primate B lymphocytes which are largely non-permissive for EBV replication (1). Further, recovery of EBV depends on its ability to growth transform normal human B lymphocytes (2). Given these limitations, mutations which affect transformation are particularly difficult to recover and characterize. Procedures for recovering mutant EBV strains irrespective of their capacity for lymphocyte growth transformation would facilitate further genetic analysis of transformation by EBV. EBV negative B lymphoma cells could be useful for virus recovery since they do not depend on EBV for their growth and are susceptible to EBV infection (3-6). In this work we investigated whether dominantly selectable drug resistance genes could be inserted into the EBV genome and used to retrieve recombinants following infection of EBV negative B lymphoma cells.