Expression of the green fluorescent protein-encoding gene from a tobacco mosaic virus-based vector

Abstract

A cDNA fragment encompassing the Aequorea victoria green fluorescent protein-encoding gene (gfp) was introduced into a genomic cDNA clone of tobacco mosaic virus (TMV). Infectious RNA transcripts produced in vitro were used to inoculate tobacco (Nicotiana benthamiana) leaves. After 1-2 days, bright green fluorescent areas could be visualized upon illumination with a long-wave ultraviolet (UV) light source. The virus was capable of infecting and expressing gfp in plant tissues both locally (in the inoculated leaf) and systemically (throughout the entire plant). Continued observation of inoculated plants indicated that systemic infection resulted from a combination of two forms of virus movement: a slow, cell-to-cell spread with concomitant virus replication and gfp expression, and a rapid, vascular-mediated form of transport without gfp expression.