Surface displayed expression of a neutralizing epitope of spike protein from a Korean strain of porcine epidemic diarrhea virus |
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Authors: | Seung-Moon Park Ae-Young Mo Jung-Gu Lim Hea-Jong Chung Tae-Geum Kim Kang-Ju Kim Dong-Ha Cho Moon-Sik Yang Dae-Hyuk Kim |
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Institution: | (1) Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University, 561-756 Jeonju, Korea;(2) Department of Oral Microbiology, School of Dentistry, Wonkwang University, 570-749 Iksan, Korea;(3) Department of Plant Biotechnology, College of Bioscience and Biotechnology, Kangwon National University, 200-701 Chuncheon, Korea |
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Abstract: | The neutralizing epitope (K-COE) of the spike protein from a Korean strain of porcine epidemic diarrhea virus (PEDV) has been
shown to prevent and foster an immune response to PED, when orally adjusted. The cell surface of the budding yeast,Saccharomyces cerevisiae, was engineered to anchor the K-COE on the outer layer of the cell, and consequently, the altered yeast was applied as a
dietary complement for animal feed, with immunogenic functions. In this study, the K-COE gene (K-COE) of the Korean strain of PEDV with the signal peptide of rice amylase 1A (Ramy 1A), was fused with the gene encoding the carboxyterminal half (320 amino acid residues from the C terminus) of yeast α-agglutinin,
a mating associated protein that is anchored covalently to the cell wall. The glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter was selected in order to direct the expression of the fusion construct, and the resulting recombinant plasmid was
then introduced intoS. cerevisiae. The surface display of K-COE was visualized via confocal microscopy using a polyclonal antibody against K-COE as the primary
antibody, and FITC (fluorescein isothiocyanate)-conjugated goat anti-mouse IgG as the secondary antibody. The display of the
K-COE on the cell surface was further verified via Western blot analysis using the cell wall fraction after the administration
of α-1,3-glucanase/PNGase F/β-mannosidase treatment. |
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Keywords: | surface display porcine epidemic diarrhea virus Saccharomyces cerevisiae |
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