The complementary-strand transcripts from Epstein-Barr virus BamHI-A-I fragments (also called the BARF0 RNAs) were first found in a nasopharyngeal carcinoma passaged in nude mice, c15, and soon proved to exist in various EBV-associated tumors and virus-carrying B cell lines. The data from previous studies revealed that the BARF0 transcripts have a highly spliced structure, whereas the evidence about their in vivo translational products remains few. The BARF0 RNA is also somehow considered as a new member of the viral latent transcripts. In order to know further about its transcriptional profile, we investigated its transcription in several EBV-carrying B cell lines during different viral stages. Our results show that (1) at least some of the BARF0 transcripts are transcribed during the viral lytic stage in the tested inducible lines; (2) the major lytic BARF0 mRNA appears heterogeneous in size (2.5 to 4.2 kb) among different lines; (3) in P3HR1 and Akata lines, the major BARF0 transcript is highly spliced and sensitive to both an inhibitor of the viral DNA polymerase and of protein synthesis. The identification of such lytic BARF0 RNAs may help us to understand further about their complex gene expression.