EFFICIENT RESISTANCE TO POTATO VIRUS Y INFECTION CONFERRED BY CYTOSOLIC EXPRESSION OF ANTI-VIRAL PROTEASE SINGLE-CHAIN VARIABLE FRAGMENT ANTIBODY IN TRANSGENIC POTATO PLANTS

M. Ayadi, D. Bouaziz, O. Nouri-Ellouz, S. Rouis, N. Drira, R. Gargouri-Bouzid

Abstract


The production of transgenic potato plants that express anti-Potato virus Y (PVY) single chain variable fragment (scFv) antibodies directed against a viral protease is reported. Complementary DNA encoding the recombinant anti-PVY NIa protease antibody was transferred to potato plants (cvs Belle de Fontenay, Claustar and Nicola) and inserted downstream of the Tobacco mosaic virus (TMV) O leader sequence in order to increase translation efficiency. Protein expression in transgenic plants was verified by Western blot and its subcellular localization by immunocytochemical analyses. The intracellular accumulation of scFv in protoplasts from transgenic potato leaves was assessed. Transgenic potato plants were transferred to a greenhouse and mechanically inoculated with PVYO. Transgenic potato plants expressing anti-PVY protease scFv antibodies in the cytosol led to complete resistance against the target virus.

Keywords


PVYO;PVY NIa protease;immunocytochemistry;recombinant antibody

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DOI: http://dx.doi.org/10.4454/JPP.FA.2012.057

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