title: Synthesis and assembly of Escherichia coli heat-labile enterotoxin B subunit in transgenic lettuce (Lactuca sativa).
abstract: Escherichia coli heat-labile enterotoxin B subunit (LTB) strongly induces immune responses and can be used as an adjuvant for co-administered antigens. Synthetic LTB (sLTB) based on optimal codon usage by plants was introduced into lettuce cells (Lactuca sativa) by Agrobacterium tumefaciens-mediated transformation methods. The sLTB gene was detected in the genomic DNA of transgenic lettuce leaf cells by PCR DNA amplification. Synthesis and assembly of the sLTB protein into oligomeric structures of pentameric size was observed in transgenic plant extracts using Western blot analysis. The binding of sLTB pentamers to intestinal epithelial cell membrane glycolipid receptors was confirmed by G(M1)-ganglioside enzyme-linked immunosorbent assay (G(M1)-ELISA). Based on the results of ELISA, sLTB protein comprised approximately 1.0-2.0% of total soluble protein in transgenic lettuce leaf tissues. The synthesis and assembly of sLTB monomers into biologically active oligomers in transgenic lettuce leaf tissues demonstrates the feasibility of the use of edible plant-based vaccines consumed in the form of raw plant materials to induce mucosal immunity.
authors.affiliation: Division of Biological Sciences and Research Center for Bioactive Materials, Chonbuk National University, Jeonju 561-756, Republic of Korea.
authors.forename: Tae-Geum
authors.initials: TG
authors.lastname: Kim
authors.forename: Mi-Young
authors.initials: MY
authors.lastname: Kim
authors.forename: Bang-Geul
authors.initials: BG
authors.lastname: Kim
authors.forename: Tae-Jin
authors.initials: TJ
authors.lastname: Kang
authors.forename: Young-Sook
authors.initials: YS
authors.lastname: Kim
authors.forename: Yong-Suk
authors.initials: YS
authors.lastname: Jang
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Arntzen
authors.forename: Moon-Sik
authors.initials: MS
authors.lastname: Yang
mesh_terms: D001427:Bacterial Toxins / Q000096:biosynthesis*; D015152:Blotting, Northern; D004768:Enterotoxins / Q000096:biosynthesis*; D029968:Escherichia coli Proteins / Q000096:biosynthesis*; D005677:G(M1) Ganglioside / Q000378:metabolism; D018545:Lactuca / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D005677:G(M1) Ganglioside; C027538:heat-labile enterotoxin, E coli