Plants are low-cost bioreactors for the production of various biopharmaceuticals including oral vaccines. Plant-derived oral
vaccines are potentially useful in combating viral infections involving mucosal immunity. Transgenic plants have been generated
to successfully produce mucosal vaccines against cholera, hepatitis B, foot-and-mouth disease, and Norwalk virus. As a first
step toward the generation of oral vaccines against the severe acute respiratory syndrome coronavirus (SARS-CoV), we have
expressed a recombinant S1 protein of the SARS-CoV in transformed tobacco. Since plant transformation and regeneration of
stable transformants require considerable time, we initially used a green fluorescent protein (GFP) to tag the antigen in
transient expression. GFP was fused to the carboxy-terminus of S1 for expression of S1-GFP to show expression of recombinant
S1 by agroinfiltration of tobacco leaves. The GFP tag enables a relatively quick confirmation of antigen expression in plant
cells by fluorescent microscopy. Such analysis using GFP that precedes stable plant transformation will enable the rapid screening
of multiple constructs to attain optimal recombinant protein expression. Furthermore, this approach determines the subcellular
localization of the recombinant protein in plant cells, providing information on optimal subcellular targeting for production
in plant bioreactors.
Keywords Autofluorescent protein tag – Plant bioreactors – Plant transformation – Recombinant subunit vaccines – Severe acute respiratory syndrome coronavirus – Subcellular localization