Noonan B, Enzmann P J, Trust T J
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
Appl Environ Microbiol. 1995 Oct;61(10):3586-91. doi: 10.1128/aem.61.10.3586-3591.1995.
Fragments of the glycoprotein genes of viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) were cloned into a bacterial broad-host-range expression vector under the control of the plac promoter. Western blot (immunoblot) analysis with monoclonal antibodies specific to the glycoproteins demonstrated the inducible expression of the fusion proteins in Escherichia coli. Aeromonas salmonicida is the causative agent of furunculosis in salmonid fish. It was confirmed that an avirulent strain of A. salmonicida, A440, which contains a deletion in the structural gene for the paracrystalline surface protein array, will provide protective immunity against furunculosis when used as a live attenuated vaccine. The plasmid-encoded viral epitopes were then mobilized into A440 for use as a shuttle system for the expression of fragments of the glycoprotein genes of IHNV and VHSV. Vaccination of rainbow trout with A440 containing the viral epitopes resulted in the development of protective immunity against both VHSV and IHNV. This indicates that the use of cloned fragments of the glycoproteins and the use of A. salmonicida as a shuttle system constitute a feasible approach to fish vaccine development.
将病毒性出血性败血症病毒(VHSV)和传染性造血器官坏死病毒(IHNV)的糖蛋白基因片段克隆到受plac启动子控制的细菌广宿主范围表达载体中。用针对糖蛋白的单克隆抗体进行的蛋白质免疫印迹(免疫印迹)分析表明,融合蛋白在大肠杆菌中可诱导表达。杀鲑气单胞菌是鲑科鱼类疖疮病的病原体。已证实,杀鲑气单胞菌无毒株A440的对晶状表面蛋白阵列结构基因有缺失,用作减毒活疫苗时可提供针对疖疮病的保护性免疫。然后将质粒编码的病毒表位导入A440,用作表达IHNV和VHSV糖蛋白基因片段的穿梭系统。用含有病毒表位的A440对虹鳟进行疫苗接种,可产生针对VHSV和IHNV的保护性免疫。这表明使用糖蛋白的克隆片段以及使用杀鲑气单胞菌作为穿梭系统是鱼类疫苗开发的一种可行方法。