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采用基因工程的方法,将副溶血弧菌的热稳定直接溶血素基因tdh2和鳗弧菌外膜蛋白基因ompU进行融合,在大肠杆菌中得到表达,并利用该融合基因构建二联DNA疫苗pEGFP-N1/tdh2-ompU。用DNA疫苗按10和50μg/尾的剂量通过肌肉注射免疫大菱鲆,对大菱鲆抵抗致病性副溶血弧菌和鳗弧菌的免疫效果进行研究。结果表明,DNA疫苗免疫的大菱鲆对副溶血弧菌感染的最高保护率为100%,对鳗弧菌感染的保护率为35%。被免疫的大菱鲆肌肉组织中能检测到融合蛋白表达,在血清中能检测到较高水平特异性抗体,DNA疫苗引起的体液免疫反应水平和保护效果与注射剂量有关。
The genetically engineered method was used to fuse the thermostable direct hemolysin gene tdh2 of Vibrio parahaemolyticus with the outer membrane protein gene ompU of Vibrio anguillarum and obtain the expression in E. coli and to construct a binary DNA vaccine pEGFP- N1 / tdh2-ompU. Immunization of turbot with intramuscular injection of DNA vaccine at doses of 10 and 50 μg / tail was carried out to study the immune effect of turbot against pathogenic V. parahaemolyticus and V. anguillarum. The results showed that the highest protection rate of V. parahaemolyticus infection by DNA vaccine-immunized turbot was 100%, and the protection against Vibrio anguillarum infection was 35%. Immunized turbot turbid muscle tissue can detect the expression of fusion protein in serum can detect higher levels of specific antibodies, DNA vaccine caused humoral immune response and the protective effect of the injection dose.