title: Assembly of cholera toxin B subunit full-length rotavirus NSP4 fusion protein oligomers in transgenic potato.
abstract: A CTB-NSP4(175) fusion gene encoding the entire 175-aa murine rotavirus NSP4 enterotoxin protein was transferred into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation. The CTB-NSP4(175) enterotoxin fusion gene was detected in the genomic DNA of transformed leaves by PCR DNA amplification. Synthesis and assembly of the full-length CTB-NSP4(175) fusion protein into oligomeric structures of pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding of CTB-NSP4(175 )fusion protein pentamers to intestinal epithelial cell membrane receptors was quantified by G(M1)-ganglioside enzyme-linked immunosorbent assay (G(M1)-ELISA). The ELISA results showed that CTB-NSP4(175) fusion protein was 0.006-0.026% of the total soluble tuber protein. The synthesis of CTB-NSP4(175) monomers and their assembly into biologically active oligomers in transformed potato tubers demonstrates the feasibility of using edible plants for the synthesis of enterocyte-targeted full-length rotavirus enterotoxin antigens that retain all of their pathogenic epitopes for initiation of a maximum mucosal immune response.
authors.affiliation: Department of Biochemistry and Microbiology and Center for Molecular Biology and Gene Therapy, School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA.
authors.forename: T-G
authors.initials: TG
authors.lastname: Kim
authors.forename: W H R
authors.initials: WH
authors.lastname: Langridge
mesh_terms: D001483:Base Sequence; D002772:Cholera Toxin / Q000235:genetics*; D017931:DNA Primers; D030821:Plants, Genetically Modified / Q000235:genetics*; D011198:Solanum tuberosum / Q000235:genetics*
chemical_list: D017931:DNA Primers; D002772:Cholera Toxin