赤点石斑鱼神经坏死病毒MCP基因原核表达条件优化

Optimization for Prokaryotic Expression of MCP Gene of Red-spotted Grouper Nervous Necrosis Virus

[目的]优化赤点石斑鱼神经坏死病毒的主衣壳蛋白(MCP)基因的原核表达条件。[方法]采用RT-PCR技术扩增出赤点石斑鱼神经坏死病毒MCP基因,构建重组表达载体pRSETA-MCP,以其转化大肠杆菌BL21(DE3)plysS,在不同培养基、温度、pH值条件下进行诱导表达。[结果]重组菌在SOB和LB培养基、pH值7.0、37℃条件下表达量最高,所得的融合蛋白分子量约为44.5 kD。[结论]该研究为RGNNV-MCP疫苗研制奠定了基础。

[ Objective] To optimize the prokaryotic expression of MCP gene of Red-spotted Grouper Nervous Necrosis Virus. [ Method] The MCP gene was amplified from Red-spotted Grouper Nervous Necrosis viral genome by RT-PCR. The recombinant expression vector pRSET A-MCP was constructed and transformed into BI21 （DE3）plysS to express proteins with induction in different media, at different pH, or at dif- ferent temperatures. [ Result ] The expression level of recombinant bacteria reached a peak with induction under the following condition： SOB or LB medium, pH 7.0, 37 ℃ while the fusion protein was about 44.5 kD in molecular weight. [ Conclusion] This study provided a basis for the development of RGNNV-MCP vaccine.