A stable cytosolic expression of VH antibody fragment directed against PVY NIa protein in transgenic potato plant confers partial protection against the virus.
The expression of recombinant antibodies in transgenic plants has been proved to be an efficient approach for large-scale production. However, the stability of these molecules and their accumulation level depend on their molecular properties and cellular targeting. The expression of single-domain antibody fragment (VH) can be advantageous since it offers small length, high expression, solubility and stability. It can therefore be preferred to other antibody derivatives avoiding the expression difficulties related to immunoglobulin domain folding via the formation of disulfide bridge. This report describes the production of transgenic potato plants expressing a VH antibody directed against the NIa protease of potato virus Y. The antibody was driven by the constitutive CaMV 35S RNA promoter. The expression cassette was transferred into potato plants via Agrobacterium tumefaciens mediated transformation. All transgenic lines showed detectable levels of VH protein confirming the efficient translation and stability of this protein. The cellular localisation of the VH antibody was investigated. Transgenic and control plants were transferred in the greenhouse and mechanically inoculated by PVY(o) suspension. Some of the transgenic lines showed delayed symptoms at the first period post inoculation and then displayed a recovery phenomenon while the virions were still detected in the leaves.
Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Ecole Nationale d'Ingénieurs de Sfax, BP 1173, 3038 Sfax, Tunisia.
Donia
D
Bouaziz
Centre de Biotechnologie de Sfax, BP, K, 3038 Sfax, Tunisia.
Malika
M
Ayadi
Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Ecole Nationale d'Ingénieurs de Sfax, BP 1173, 3038 Sfax, Tunisia.
Amira
A
Bidani
Centre de Biotechnologie de Sfax, BP, K, 3038 Sfax, Tunisia.
Souad
S
Rouis
Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Ecole Nationale d'Ingénieurs de Sfax, BP 1173, 3038 Sfax, Tunisia; Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Institut Préparatoire aux Etudes d'Ingénieurs, BP 1172, 3018 Sfax, Tunisia.
Oumèma
O
Nouri-Ellouz
Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Ecole Nationale d'Ingénieurs de Sfax, BP 1173, 3038 Sfax, Tunisia.
Raïda
R
Jellouli
Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Ecole Nationale d'Ingénieurs de Sfax, BP 1173, 3038 Sfax, Tunisia; Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Faculté des Sciences de Sfax, BP 1171, 3038 Sfax, Tunisia.
Noureddine
N
Drira
Laboratoire des Biotechnologies Végétales Appliquées à L'Amélioration des Cultures, Ecole Nationale d'Ingénieurs de Sfax, BP 1173, 3038 Sfax, Tunisia. Electronic address: gargouriradhia@yahoo.fr.
Radhia
R
Gargouri-Bouzid
NIa protease; PVY; Recombinant VH antibody; Solanum tuberosum; Transgenic plants