The Borna disease virus antigenome includes five major open reading frames (ORFs) which encode, from 5' to 3', the putative nucleoprotein (N), the phosphoprotein (P), the putative matrix protein (M), the major glycoprotein (G), and the RNA-dependent RNA polymerase (pol). Whereas the N and P ORFs are translated from monocistronic transcripts, the M, G, and pol ORFs are translated from polycistronic transcripts. Expression of the M, G, and pol ORFs is dependent upon differential splicing of two introns (intron 1, 94 nucleotides [nt]; intron 2, 1,294 nt). In vitro transcription-translation assays of wild-type and mutant sequences indicated that the G ORF is translated from an unspliced 2.8-kb RNA by leaky ribosomal scanning. Splicing of intron 1 enhances the translation of the G ORF by converting the M ORF into a 13-amino-acid minicistron, a structure that facilitates ribosomal reinitiation.