Production of double repeated B subunit of Shiga toxin 2e at high levels in transgenic lettuce plants as vaccine material for porcine edema disease

• Published in: Transgenic research Vol. 20; no. 4; pp. 735 - 748
• Main Authors: Matsui, Takeshi, Takita, Eiji, Sato, Toshio, Aizawa, Michie, Ki, Misa, Kadoyama, Yumiko, Hirano, Kenji, Kinjo, Satoko, Asao, Hiroshi, Kawamoto, Keiko, Kariya, Haruko, Makino, Sou-Ichi, Hamabata, Takashi, Sawada, Kazutoshi, Kato, Ko
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.08.2011; Springer; Springer Nature B.V
• Subjects: Animal Genetics and Genomics; Animals; Bacterial Vaccines - genetics; Bacterial Vaccines - therapeutic use; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology; Edema Disease of Swine - therapy; Enterohemorrhagic Escherichia coli; Escherichia coli; Fundamental and applied biological sciences. Psychology; Genetic Engineering; Genetic technics; Genetic Vectors; Lactuca - genetics; Lactuca - metabolism; Life Sciences; Methods. Procedures. Technologies; Molecular Medicine; Original Paper; Plant Genetics and Genomics; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Protein Subunits - genetics; Shiga Toxin 2 - biosynthesis; Shiga Toxin 2 - genetics; Shiga Toxin 2 - therapeutic use; Swine; Transgenic animals and transgenic plants; Transgenics; Vaccines, Edible - genetics; Vaccines, Edible - therapeutic use; Edible vaccine; Lettuce; Shiga toxin 2e B; Pig edema disease; Edema; Disease; Materials; Shiga toxin 2e B; Vaccine; Subunit; Pig; Vertebrata; Mammalia; Production; Transgenic plant; Artiodactyla; Ungulata; Shiga like toxin
• ISSN: 0962-8819; 1573-9368
• DOI: 10.1007/s11248-010-9455-9

Pig edema disease is a bacterial disease caused by enterohemorrhagic Escherichia coli . E. coli produces Shiga toxin 2e (Stx2e), which is composed of one A subunit (Stx2eA) and five B subunits (Stx2eB). We previously reported production of Stx2eB in lettuce plants as a potential edible vaccine (Matsui et al. in Biosci Biotechnol Biochem 73:1628–1634, 2009 ). However, the accumulation level was very low, and it was necessary to improve expression of Stx2eB for potential use of this plant-based vaccine. Therefore, in this study, we optimized the Stx2eB expression cassette and found that a double repeated Stx2eB (2× Stx2eB) accumulates to higher levels than a single Stx2eB in cultured tobacco cells. Furthermore, a linker peptide between the two Stx2eB moieties played an important role in maximizing the effects of the double repeat. Finally, we generated transgenic lettuce plants expressing 2× Stx2eB with a suitable linker peptide that accumulate as much as 80 mg per 100 g fresh weight, a level that will allow us to use these transgenic lettuce plants practically to generate vaccine material.