Expression and Immunogenicity of the Mycobacterial Ag85B/ESAT-6 Antigens Produced in Transgenic Plants
by Elastin-Like Peptide Fusion Strategy
Production of plant-expressed TBAg-ELP fusion protein. (a) Diagrammatic representation of the TBAg-ELP expression cassette. Expression of the recombinant fusion protein was driven by the constitutive CaMV 35S promoter. Endoplasmic reticulum retrieval was mediated by a C-terminal KDEL tag and the c-myc tag was included to facilitate detection. ELP fusion consisted of 100 repeats of the pentapeptide VPGXG, where X was valine, glycine, or alanine (100xELP). (b) Accumulation increase of TBAg-ELP in transgenic tobacco plants. Western blot analysis of leaf extracts of individual plants expressing the transgene. Proteins were separated by reducing 10% SDS-PAGE, blotted, and immunodetected using an anti-c-myc antibody. Numbers designate the independent plants. The same amounts of TSP were loaded on the gel (15 g/lane). S: cmyc standard (15 ng/lane); Wt: wild-type N. tabacum.