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Transgenic Res ; 11(5): 447-54, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12437076

ABSTRACT

Cholera toxin, secreted by Vibrio cholerae, consists of A and B subunits. The latter binds to G(M1)-ganglioside receptors as a pentamer (approximately 55 kDa). Tomato plants were transformed with the gene encoding cholera toxin B subunit (ctxB) along with an endoplasmic reticulum retention signal (SEKDEL) under the control of the CaMV 35S promoter via Agrobacterium-mediated transformation. PCR and Southern analysis confirmed the presence of the ctxB gene in transformed tomato plants. Northern analysis showed the presence of the ctxB-specific transcript. Immunoblot assays of the plant-derived protein extract showed the presence of cholera toxin subunit B (CTB) with mobility similar to purified CTB from V. cholerae. Both tomato leaves and fruits expressed CTB at levels up to 0.02 and 0.04% of total soluble protein, respectively. The G(M1)-ELISA showed that the plant-derived CTB bound specifically to G(M1)-ganglioside receptor, suggesting that it retained its native pentameric form. This study forms a basis for exploring the utility of CTB to develop tomato-based edible vaccines against cholera.


Subject(s)
Agrobacterium tumefaciens/genetics , Cholera Toxin/genetics , Plants, Genetically Modified , Solanum lycopersicum/genetics , Blotting, Northern , Blotting, Southern , Blotting, Western , Cholera Toxin/metabolism , DNA Primers/chemistry , Endoplasmic Reticulum/metabolism , Enzyme-Linked Immunosorbent Assay , G(M1) Ganglioside/metabolism , Gene Expression Regulation, Plant , Gene Transfer Techniques , Genetic Vectors , Solanum lycopersicum/metabolism , Plant Leaves/chemistry , Plasmids , Polymerase Chain Reaction , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, Cell Surface/metabolism
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