Infection of the human T-cell line MT-2 by Epstein-Barr virus(EBV) recombinants with a hygromycin resistance gene and subsequent selection with this drug permitted isolation and long-term maintenance of EBV-infected MT-2 clones. These MT-2 clones harbor monoclonal episomes of EBV DNA and stably express two EBV latent proteins, EBV nuclear antigen 1(EBNA1) and latent membrane protein 1 (LMP1). Some MT-2 clones also express EBNA3C, yet none of the clones express EBNA2. This pattern of EBV gene expression is similar to that observed in most cases of EBV-positive peripheral T-cell lymphoma. In addition to these latent viral functions, expression of some early EBV replicative genes, including BZLF1, was also detected. EBV recombinants with positive selection markers will be a useful tool in establishing experimental systems of persistent EBV infection in certain non-B-cell lineages.