Transcription and DNA sequence of the BamHI L fragment of B95-8 Epstein-Barr virus

Abstract

The sequence of the BamHI L fragment of B95-8 Epstein-Barr virus (EBV) DNA has been determined. The transcription starts of five promoters have been mapped to this fragment, using S1 mapping and either in vitro transcription or the primer extension technique. Dramatically increased levels of cytoplasmic poly(A) + RNAs, transcribed from these promoters, occur after treatment of B95-8 cells with 12-O-tetradecanoylphorbol-13-acetate (TPA). For three of these promoters addition of phosphonoacetic acid (PAA) inhibits the effect of TPA, indicating that they give rise to late lytic cycle RNAs. The other two promoters give rise to early RNAs. Northern blot analysis indicates that one of the late promoters initiates two transcripts whose size differences are due to different splicing patterns. These two RNAs code for the 350/300 and 250/200 kd envelope glycoproteins of EBV. The sequences of these proteins would be of use in the production of a synthetic vaccine to prevent EBV infection.