HE Wen-bin 1, 2, 3, XU Li-ming2，ZHAO Jing-zhuang2, LIU Miao2, REN Guang-ming2, LU Tong-yan2, YIN Jia-sheng2*
1.National Demonstration Centre for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China； 2.Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070,China； 3.Aquatic Pathogen Collection Center，Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China
Abstract: The VP2 gene， as the most important surface antigen of infectious pancreatic necrosis (IPN), was amplified by RT-PCR with specific PCR primers based on IPNV ChRtm213 VP2 gene, and the PCR product was ligated to pYD1 vector and the reliable plasmid was named as pYD1-VP2. The Saccharomycescerevisiae strain EBY100 was transformed with pYD1-VP2 plasmid using electroporation, and then induced by galactose to express VP2 protein on yeast cell surface. The Western Blotting and immunofluorescence assay revealed that VP2 expression and a specific VP2 protein were observed in the induced EBY100/pYD1-VP2 cell which presented a specific green fluorescence on the cell surface, with positive correlated relationship between fluorescence intensity and induction time, with significant differences in proportion of fluorescent yeast among 24 h, 36 h, and 48 h, and control groups. The findings indicated that the VP2 protein was successfully displayed on the yeast cell surfaces and that provided the basis with oral living vector vaccine of IPNV.
2019, Vol. 34 
