Transient expression of an active human interferon-beta in lettuce

Using Agrobacterium mediated transient expression method, plant bivalent expression vector pBI121 containing GUS as a report gene was transformed into lettuce (Lactuca sativa). Through designed orthogonal analysis, intact lettuce leaves infiltrated with 200 mu M acetosyringone and 0.8 OD600 bacterial suspensions under vacuum for 30 min, then co-cultured at 24 degrees C for 6 ds produced a maximum GUS protein of 2.5% TSP with 21.39 nmol mg(-1) min(-1) MU activity, which was 19 times of the control (1.31 mnol mg(-1) min(-1) MU). Employed these optimized conditions HuIFN-beta was expressed in lettuce leaves. Western blot and antivirus bioactivity analyses confirmed the HuIFN-beta achieved by agrobacterium infiltration had a high biological activity (3.1 x 10(4) IU/mL). To our knowledge, it is the first detailed orthogonal optimizing study of Agrobacterium, mediated transient expression and the first report on the production of the biologically active therapeutic proteins produced by Agrobacterium mediated transient expression in lettuce. In summary, transient expression by Agrobacterium vacuum infiltration can be adopted as an efficient, inexpensive and small-scaled plant expression system for therapeutic protein production. (c) 2006 Elsevier B.V. All rights reserved.