Foamy virus (FV) is an unconventional retrovirus that possesses a complex genome and a special mechanism for gene expression regulation. The genome encodes transcriptional protein Tas which is found to regulate both the internal promoter (IP) and the long terminal repeat promoter (LTR). However, the detailed mechanism of Tas-mediated gene expression remains unknown. In this study, we provided the first evidence for the temporal production and utilization of four different bovine foamy virus (BFV) btas mRNAs during persistent infection. These four forms of btas mRNA transcripts initiated either at BFV LTR or IP and spliced or unspliced have a differential ability to activate BFV promoters. Furthermore, by developing an MS2 translational operator/coat protein combined system to track mRNA exportation from the nucleus and distribution throughout the cytoplasm, we observed that the IP spliced transcript could be exported into the cytoplasm more efficiently than unspliced transcripts. These findings provide evidence for the hypothesis that the functional interplay of both promoters contributes to the temporal pattern of BFV transcription and suggest that a post-transcriptional regulation exist in BFV replication.