Synthesis and assembly of a cholera toxin B subunit-rotavirus VP7 fusion protein in transgenic potato |
| |
Authors: | Nak-Won Choi Mary K Estes William H R Langridge |
| |
Institution: | (1) Center for Molecular Biology and Gene Therapy, Department of Biochemistry and Microbiology, School of Medicine, Loma Linda University, 92354 Loma Linda, CA;(2) Department of Molecular Virology and Microbiology, Baylor College of Medicine, 77030 Houston, Texas |
| |
Abstract: | A gene encoding VP7, the outer capsid protein of simian rotavirus SA11, was fused to the carboxyl terminus of the cholera
toxin B subunit gene. A plant expression vector containing the fusion gene under control of the mannopine synthase P2 promoter was introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation. The CTB::VP7 fusion gene was detected in the genomic DNA of transformed potato leaf cells by polymerase chain reaction (PCR) amplification
methods. Immunoblot analysis of transformed potato tuber tissue extracts showed that synthesis and assembly of the CTB::VP7
fusion protein into oligomers of pentameric size occurred in the transformed plant cells. The binding of CTB::VP7 fusion protein
pentamers to sialo-sugar containing GM1 ganglioside receptors on the intestinal epithelial cell membrane was quantified by
enzyme-linked immunosorbent assay (ELISA). The ELISA results showed that the CTB::VP7 fusion protein made up approx 0.01%
of the total soluble tuber protein. Synthesis and assembly of CTB::VP7 monomers into biologically active pentamers in transformed
potato tubers demonstrates the feasibility of using edible plants as a mucosal vaccine for the production and delivery system
for rotavirus capsid protein antigens. |
| |
Keywords: | Rotavirus cholera toxin edible vaccine adjuvant Solanum tuberosum |
本文献已被 PubMed SpringerLink 等数据库收录! |