Expression of Helenium virus S coat protein in Escherichia coli, in vitro in rabbit reticulocyte lysate and transgenic tobacco.

The coat protein open reading frame (ORF) sequence of Helenium virus S (HelVS) was cloned and expressed in E. coli, rabbit reticulocyte and transgenic tobacco. In E. coli the size of the protein was identical to that obtained for the coat protein from purified virus particles and less than that predicted for the fusion protein. This may be due to ribosome binding at a potential ribosome binding site present on the viral sequence, approximately 45 nucleotides upstream from the initiating methionine of the coat protein ORF. This region of HelVS, equivalent to the 1.5 kb subgenomic RNA, also produced high levels of protein when transcribed and translated in vitro. When introduced into Nicotiana tabacum by leaf disk transformation via Agrobacterium tumefaciens, high levels of stable coat protein were detected which were identical in molecular weight to that of HelVS coat protein and constituted approximately 0.1-0.5% of the total extracted protein.