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.
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 to particular domains on the cell surface. For use in bioindustrial processes applied to oral vaccination, we consider that cell-surface display systems must be useful and that the yeast Saccharomyces cerevisiae, the most suitable microorganism for practical purposes, is available as a host for genetic engineering because it can be subjected to many genetic manipulations. In particular, 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 sea bream iridovirus (RSIV), which is one of the most common viral diseases in the cultured marine fish Pagrus major in Japan, using the arming yeast system and aiming at its application for oral vaccination. We first performed the molecular cloning and expression of the 380R antigen from RSIV in Escherichia coli. The nucleotide sequence of the 380R antigen was composed of an open reading frame (ORF) of 1360 bp encoding a protein of 453 residues. To prepare a specific antibody against the 380R antigen, the recombinant protein was overexpressed and purified in E. coli. As a result of indirect immunofluorescence with the specific antibody, we could observe the expression of the 380R antigen on the surface of the yeast cells. Thus, we have successfully prepared the source of an oral vaccine using cell-surface display technology in yeast.
Tamaru
Yutaka
Y
Department of Life Science, Graduate School of Bioresources, Mie University, Tsu, Japan. ytamaru@bio.mie-u.ac.jp
Ohtsuka
Masanori
M
Kato
Keitaro
K
Manabe
Sadao
S
Kuroda
Kouichi
K
Sanada
Mitsuaki
M
Ueda
Mitsuyoshi
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D018014:Gene Transfer Techniques; D017994:Iridovirus / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D021541:Sea Bream / Q000821:virology*; D014612:Vaccines / Q000737:chemistry* / Q000302:isolation & purification
D036022:Capsid Proteins; D004279:DNA, Viral; D014612:Vaccines