Figure 13.

INPACT-Directed Expression of Human Vitronectin in Transgenic N. benthamiana.

(A) Time course of Vitronectin expression in detached leaves of wild-type (WT) and transgenic plants containing the INPACT-hVN cassette (NbSRN2 + INPACT-hVN) activated in a Murashige and Skoog suspension containing 0.5% ethanol. Samples were taken at days 0, 2, 4, and 6 PEA. TSP was separated by PAGE and analyzed by immunoblotting with an anti-human Vitronectin monoclonal antibody (The Antibody Shop). A positive control lane was included containing 100 ng of purified human Vitronectin (Promega).

(B) Time course of Vitronectin expression in soil-acclimated transgenic plants containing the INPACT-hVN cassette. Samples were taken from the top and bottom leaves over a 5-d period (0 to 5) following activation with a 1% ethanol solution and analyzed by immunoblotting.

(C) and (D) Coomassie blue–stained PAGE (C) and immunoblot analysis (D) tracking partial purification of plant-made Vitronectin using heparin sepharose and HIS-tag affinity columns. M, prestained SDS-PAGE standards, low range (Bio-Rad); FT, flow-through; W, wash; E, elution (E1-5 represent fractions 1 to 5).