Identification and characterization of recombinant subgroup J avian leukosis viruses (ALV) expressing subgroup A ALV envelope

Three recombinant avian leukosis subgroup J viruses, ADOL 5701A, ADOL 5701A Delta, and ADOL 6803A, carrying a subgroup A envelope have been isolated and characterized. These viruses were identified by their ability to replicate in DF-1/J, a recombinant chicken embryo fibroblast (CEF) cell line expressing the subgroup J envelope that is resistant to subgroup J replication. Flow cytometric analysis of DF-1/J cells infected with ADOL 5701 and ADOL 6803, two subgroup J isolates, indicated cross-reactivity with subgroup A chicken polyclonal serum. Based on published sequences of subgroups A and J isolates, we designed a series of primers to PCR amplify the envelope and LTR of these viruses. PCR products were obtained when the forward primer was specific for subgroup A gp85 envelope protein gene and the reverse primer was specific for subgroup J LTR. Sequence analysis of the PCR products indicated that these viruses had a subgroup A gp85, a subgroup E gp37, and a subgroup J LTR. Interestingly, these Viruses had previously been propagated in CEF from the alv6 chicken line, a line that carries a replication defective recombinant endogenous virus expressing a subgroup A envelope (RAV 0-A(1)). Sequence analysis of RAV 0-A(1) gp85 and gp37 envelope proteins indicated that they were almost identical to those of the recombinants ADOL 5701A and ADOL 6803A. These results indicate that these three recombinant viruses arose by recombination between exogenous subgroup J isolates and a recombinant defective endogenous Virus with subgroup A envelope. (C) 2000 Academic Press.