Graduation: C.C. Lacorte:"Improving and assessing viral vectors for recombinant protein production in plants"

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1 Dec 2006 16:00
Unit: Wageningen UR
Location: Aula, building 362, Gen. Foulkesweg 1, Wageningen
Organisation: Wageningen University
Promotor: prof.dr. R.W. Goldbach (Virology)
Co Promotor: Dr.ir.M.W. Prins

 This research thesis focused on the use of transient expression systems for heterologous protein production in plants. Viral vectors based on PVX and TMV were converted to be compatible with the GatewayTM cloning system facilitating the insertion of target genes. Proteins VP1, VP2 and VP3 from Chicken anemia virus (CAV) were expressed in N. benthamiana plants to be tested as a potential subunit vaccine. Expression was achieved using Agrobacterium transient assay (ATTA) and the subcellular localization of these proteins studied as GFP fusions. The expression of erythropoietin (EPO) was also evaluated in plants using the TMV and PVX-base viral vectors. To facilitate purification of heterologous proteins produced in plants, a method based on gene fusions with the nucleoprotein (N) of Tomato spotted wilt virus was also evaluated using GFP as a marker gene, resulting in the formation of large aggregates that were easily purified by simple centrifugation and filtration steps.
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