The purpose of this work has been to investigate transcription and translation products from an open reading frame (ORF) located downstream of the coat protein (CP) cistron on RNA-2 of pea early browning virus (PEBV). This work was initiated as a step towards elucidation of the significance of this putative gene. Sequence data on RNA-2 suggest that a 29600 (29·6K) protein is translated from the ORF in question. Hybridization with ORF-specific probes on Northern blots with RNA from infected plants showed that PEBV synthesizes a subgenomic RNA encoding CP (RNA-2a) with a size of 3000 nucleotides (nt) and that a putative subgenomic RNA (RNA-2b) encoding the 29·6K protein appears to have a size of 1600 nt. This is 300 nt less than the size predicted from the sequencing data. For antibody production, a cDNA fragment harbouring 85% of the 29·6K ORF was cloned into the pUEX3 expression vector. The resulting plasmid expresses the 29·6K protein as a fusion protein with β-galactosidase and this protein was used for raising antiserum containing specific anti-29·6K protein antibodies. By using these antibodies on immunoblots it was demonstrated that the 29·6K protein is expressed in infected plants.


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