Application of the Arming System for the Expression of the 380R Antigen from Red Sea Bream Iridovirus (RSIV) on the Surface of Yeast Cells: A First Step for the Development of an Oral Vaccine

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• 作者：Yutaka Tamaru ; Masanori Ohtsuka ; Keitaro Kato ; Sadao Manabe ; Kouichi Kuroda ; Mitsuaki Sanada ; Mitsuyoshi Ueda
• 刊名：Biotechnology Progress
• 出版年：2006
• 出版时间：August 2006
• 年：2006
• 卷：22
• 期：4
• 页码：949 - 953
• 全文大小：455K
• 年卷期：v.22,no.4(August 2006)
• ISSN：1520-6033

文摘

The cell surface is a functional interface between the inside and the outside of the cell. Moreover,cells have systems for anchoring surface specific proteins and for confining surface proteins toparticular domains on the cell surface. For use in bioindustrial processes applied to oralvaccination, we consider that cell-surface display systems must be useful and that the yeastSaccharomyces cerevisiae, the most suitable microorganism for practical purposes, is availableas a host for genetic engineering because it can be subjected to many genetic manipulations. Inparticular, the rigid structure of the cell makes the yeast suitable for several of the applications.In this study, we describe the expression of one of the target antigens, 380R, from the red seabream iridovirus (RSIV), which is one of the most common viral diseases in the cultured marinefish Pagrus major in Japan, using the arming yeast system and aiming at its application for oralvaccination. We first performed the molecular cloning and expression of the 380R antigen fromRSIV in Escherichia coli. The nucleotide sequence of the 380R antigen was composed of anopen reading frame (ORF) of 1360 bp encoding a protein of 453 residues. To prepare a specificantibody against the 380R antigen, the recombinant protein was overexpressed and purified inE. coli. As a result of indirect immunofluorescence with the specific antibody, we could observethe expression of the 380R antigen on the surface of the yeast cells. Thus, we have successfullyprepared the source of an oral vaccine using cell-surface display technology in yeast.