Production of Red sea bream iridovirus and viral nervous necrosis virus capsid protein by yeast surface display method

Abstract

Marine viruses are the most numerous biological entities in the oceans and probably infect all living organisms from bacteria to mammal. However, there are few researches on marine viruses to isolate and detect them because of the high mutation rates and variation of viral genomes. We are currently developing new experiment system to identify and detect various marine viruses by using yeast surface display followed by scFv way. Yeast surface display (YSD) is a powerful method for isolating and engineering proteins to increase their affinity, specificity and stability in many different areas. Recently YSD has been introduced to identify and screen a certain antibody to a specific antigen protein. Yeast has eukaryotic characteristics in protein biosynthesis so that antigen protein seems to be similar 3D structures and post translational modification. In this study we expressed the capsid proteins of two marine viruses (RSIV: Red sea bream iridovirus and VNNV: Viral Nervous Necrosis virus) in YSD. Viral capsid genes were synthesized on the basis of sequence data bases and subcloned into yeast expression vector. And then these vectors were introduced into EBY100 yeast cells. FACS analysis showed that RSIV and VNNV capsid proteins were expressed good enough to isolate as antigens. Western blot result also shows RSIV and VNNV capsid protein expressed on yeast surface. Each anti RSIV- and VNNV-scFv could select from scFv phtion rates and variation of viral genomes. We are currently developing new experiment system to identify and detect various marine viruses by using yeast surface display followed by scFv way. Yeast surface display (YSD) is a powerful method for isolating and engineering proteins to increase their affinity, specificity and stability in many different areas. Recently YSD has been introduced to identify and screen a certain antibody to a specific antigen protein. Yeast has eukaryotic characteristics in protein biosynthesis so that antigen protein seems to be similar 3D structures and post translational modification. In this study we expressed the capsid proteins of two marine viruses (RSIV: Red sea bream iridovirus and VNNV: Viral Nervous Necrosis virus) in YSD. Viral capsid genes were synthesized on the basis of sequence data bases and subcloned into yeast expression vector. And then these vectors were introduced into EBY100 yeast cells. FACS analysis showed that RSIV and VNNV capsid proteins were expressed good enough to isolate as antigens. Western blot result also shows RSIV and VNNV capsid protein expressed on yeast surface. Each anti RSIV- and VNNV-scFv could select from scFv ph