title: Expression and assembly of cholera toxin B subunit and domain III of dengue virus 2 envelope fusion protein in transgenic potatoes.
abstract: The rates of mosquito-transmitted dengue virus infection in humans have increased in tropical and sub-tropical areas. Domain III of dengue envelope protein (EDIII) is involved in cellular receptor binding and induces serotype-specific neutralizing antibodies. EDIII fused to the B subunit of Vibrio cholera (CTB-EDIII) was expressed in potatoes to develop a plant-based vaccine against dengue virus type 2. CTB-EDIII fused to an endoplasmic reticulum (ER) retention signal, SEKDEL, was introduced into potatoes by A. tumefaciens-mediated gene transformation. The integration of the CTB-EDIII fusion gene into the nuclear genome of transgenic plants was confirmed by genomic DNA polymerase chain reaction (PCR), and mRNA transcripts of CTB-EDIII were detected. CTB-EDIII fusion protein was expressed in potato tubers and assembled into a pentameric form capable of binding monosialotetrahexosylganglioside (GM1). The level of expression was determined to be ∼0.005% of total soluble protein in potato tubers. These results suggest that dengue virus antigen could be produced in potatoes, raising the possibility that edible plants are employed in mucosal vaccines for protection against dengue infection.
authors.affiliation: Department of Molecular Biology, Chonbuk National University, Jeonju, Republic of Korea.
authors.forename: Mi-Young
authors.initials: MY
authors.lastname: Kim
authors.affiliation: Department of Molecular Biology, Chonbuk National University, Jeonju, Republic of Korea.
authors.forename: Jing-Ying
authors.initials: JY
authors.lastname: Li
authors.affiliation: Research Center of Bioactive Materials, Chonbuk National University, Jeonju, Republic of Korea.
authors.forename: Nguyen-Quang-Duc
authors.initials: NQ
authors.lastname: Tien
authors.affiliation: Department of Molecular Biology, Chonbuk National University, Jeonju, Republic of Korea; Department of Bioactive Material Sciences, Chonbuk National University, Jeonju, Republic of Korea; Research Center of Bioactive Materials, Chonbuk National University, Jeonju, Republic of Korea. Electronic address: mskyang@jbnu.ac.kr.
authors.forename: Moon-Sik
authors.initials: MS
authors.lastname: Yang
mesh_terms: D002772:Cholera Toxin / Q000235:genetics / Q000378:metabolism*; D053059:Dengue Vaccines / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003716:Dengue Virus; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011198:Solanum tuberosum / Q000235:genetics / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D053059:Dengue Vaccines; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D002772:Cholera Toxin
keywords: Cholera toxin B subunit; Dengue; Plant-based vaccine; Transgenic potato