Journal of General Virology

Fig. 1. (A) Accumulation of steady-state levels of transgene mRNA in N. benthamiana plants transformed with the PVA 5´-UTR+CP construct. Total RNA was extracted from six upper leaves immediately before inoculation with PVY (samples 1–6) and from the upper leaves of the same plants (samples 1´–6´) and from a non-inoculated control plant (c) 14 days after inoculation with PVY. PVY infection of the leaves was verified by ELISA. RNA (25 µg) from each sample was subjected to Northern analysis (Sambrook et al., 1989) using ³²P-labelled PVA CP cDNA as a probe. Long exposure (48 h) was used and the signals were visualized by a phosphoimager (Molecular Dynamics). Note that distinct bands corresponding to the expected size of transgene mRNA could already be seen after 2 h of exposure in samples 1´–6´. (B) Ethidium bromide staining of ribosomal RNAs showing equal loading of the samples.

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