Subcellular location and expression of tobacco Necrosis necrovirus p7a protein

A putative movement protein (p7a) of tobacco necrosis Necrovirus, strain D (TNV-D), produced in Escherichia coli using an expression vector, was used to raise an antiserum. Immunoblot analysis using this antiserum showed that the p7a protein was detectable only in the combined cell wall and cell membrane fraction prepared from TNV-D infected Phaseolus vulgaris leaves. The p7a protein was detectable 1 day after inoculation and reached a maximum 3 days later, before declining, whereas coat protein was not detectable until 3 days after inoculation and continued to increase in concentration for a further 2 days before declining. Differences in the detectable amounts of both proteins may reflect their differential stability in extracts from necrotic tissue and/or the transient expression of the putative movement protein early in the replication cycle of TNV-D.