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Expression of Escherichia coli heat-labile enterotoxin b subunit (LTB) in carrot (Daucus carota L.) |
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| Authors: | Sergio Rosales-Mendoza Ruth Elena Soria-Guerra María Teresa de Jesús Olivera-Flores Rubén López-Revilla Gerardo Rafael Argüello-Astorga Juan Francisco Jiménez-Bremont Ramón Fernando García-de la Cruz Juan Pablo Loyola-Rodríguez Ángel Gabriel Alpuche-Solís |
| Affiliation: | (1) División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, 78216 San Luis Potosí, S.L.P., Mexico;(2) Facultad de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, 04510 México, D.F., Mexico;(3) Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava No. 6, 78210 San Luis Potosí, S.L.P., Mexico;(4) Maestría en Ciencias Odontológicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava No. 2, 78290 San Luis Potosí, S.L.P., Mexico |
| Abstract: | We expressed the B subunit of enterotoxigenic Escherichia coli heat-labile enterotoxin (LTB) encoded by a synthetic codon-optimized gene in carrot. An Agrobacterium-mediated transformation method was used. Thirty independent transgenic lines were regenerated via somatic embryogenesis after 6 months in culture and were transferred to a greenhouse. GM1-ELISA assay was used to assess LTB protein content in mature taproots. Some transgenic lines expressed LTB up to 0.3% of the total soluble protein, which is tenfold higher than the expression levels reported earlier using the native bacterial gene in plants. Immunological assay confirmed proper assembly of the pentameric complex and in vitro activity of the recombinant LTB protein, suggesting that it can be functional in prevention of diarrhea. |
| Keywords: | Diarrhea prevention Plant based-vaccine Synthetic gene Expression levels Somatic embryogenesis |
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