Expression of Escherichia coli heat-labile enterotoxin b subunit (LTB) in carrot (Daucus carota L.)

• Published in: Plant cell reports Vol. 26; no. 7; pp. 969 - 976
• Main Authors: Rosales-Mendoza, Sergio, Soria-Guerra, Ruth Elena, de Jesús Olivera-Flores, María Teresa, López-Revilla, Rubén, Argüello-Astorga, Gerardo Rafael, Jiménez-Bremont, Juan Francisco, García-de la Cruz, Ramón Fernando, Loyola-Rodríguez, Juan Pablo, Alpuche-Solís, Ángel Gabriel
• Format: Journal Article
• Language: English
• Published: Berlin Berlin/Heidelberg : Springer-Verlag 01-07-2007; Springer; Springer Nature B.V
• Subjects: Bacteria; Bacterial Toxins - genetics; Bacterial Toxins - metabolism; Biological and medical sciences; Biotechnology; Daucus; Daucus carota; Daucus carota - genetics; Daucus carota - growth & development; Daucus carota - metabolism; Diarrhea prevention; E coli; Embryonic growth stage; Enterotoxins - genetics; Enterotoxins - metabolism; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Expression levels; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant - physiology; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; Plant based-vaccine; Plants, Genetically Modified; Proteins; somatic embryogenesis; Synthetic gene; Transgenic animals and transgenic plants; Transgenic plants; Daucus carota; Escherichia coli; Diarrhea; Diarrhea prevention Plant based-vaccine Synthetic gene Expression levels Somatic embryogenesis; Vaccine; Gene expression; Subunit; Somatic embryo; Prevention; Toxin; Dicotyledones; Angiospermae; Enterotoxin; Bacteria; Spermatophyta; Transgenic plant; Umbelliferae; Enterobacteriaceae
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-007-0310-2

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.