Expression and Identification of Glycoprotein of Siniperca chuatsirhabdovirus in Baculovirus Expression System
ZHANG Yanbing1,2, YE Jiaxin2, SUN Wei2, LIU Xiaodan2, LIN Qiang1
1. Key Laboratory of Fishery Drug Development, Ministry of Agriculture and Rural Affairs, Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China; 2. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China
Abstract：Siniperca chuatsi rhabdovirus (SCRV) genome encodes five structural proteins, in which glycoprotein G is the main antigen on the surface of the virus, In order to achieve large expression of the antigen protein, the glycoprotein G of SCRV was inserted into Baculovirus shuttle vector pFastBac1 by molecular cloning technology and transformed into Escherichia coli DH5α competent cells. Then it was transformed into DH10Bac E. coli competent cells, and the recombinant plasmid rBacmid-G was obtained by blue-white spot and resistance screening. The recombinant bacmid was transfected into Sf9 insect cells by liposome transfection method to obtain recombinant Baculovirus expressing glycoprotein. The obtained P3 generation recombinant virus was infected with Sf9 insect cells for SDS-PAGE detection of the recombinant protein. After successful expression, the target protein was purified by nickel ion affinity chromatography column, and then the recombinant protein was identified by Western blot. There was a specific band at 50 ku, and no band in the control group, indicating that the prepared antigen can specifically bind to the antibody. Therefore, the SCRV glycoprotein G was successfully expressed in the Baculovirus expression system. The findings provide technical support for the development and large-scale production of SCRV glycoprotein G vaccine.
张彦冰, 叶加鑫, 孙威, 刘晓丹, 林强. 鳜弹状病毒糖蛋白在杆状病毒表达系统中的表达与鉴定[J]. 水产科学, 2024, 43(2): 281-288.
ZHANG Yanbing, YE Jiaxin, SUN Wei, LIU Xiaodan, LIN Qiang. Expression and Identification of Glycoprotein of Siniperca chuatsirhabdovirus in Baculovirus Expression System. Fisheries Science, 2024, 43(2): 281-288.
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2024, Vol. 43 
