摘要
为了探讨原核表达的坦布苏病毒E蛋白结构域Ⅰ/Ⅱ蛋白对于坦布苏病毒感染的免疫保护作用,利用IPTG诱导重组大肠杆菌表达坦布苏病毒E蛋白结构域Ⅰ/Ⅱ蛋白,Anti-FLAG M2 Affinity Gel纯化后进行SDS-PAGE和Western blot鉴定。经SDS-PAGE和Western blot鉴定,纯化后坦布苏病毒E蛋白结构域Ⅰ/Ⅱ蛋白仅有单一目的条带。免疫组小鼠血清ELISA效价均达4.67±0.11(lg10)。攻毒保护试验结果表明,免疫组小鼠体质量在攻毒9 d后高于对照组,荧光定量RT-PCR检测结果显示,攻毒后免疫组小鼠脑、肝脏、脾脏组织中坦布苏病毒核酸含量均低于对照组。表明使用原核表达的坦布苏病毒E蛋白结构域Ⅰ/Ⅱ蛋白免疫可诱导产生高水平抗体,并提供良好的免疫保护力,为开发有效的坦布苏病毒亚单位疫苗提供依据。
The immunogenicity of the prokaryotically-expressed Tembusu virus E protein domains Ⅰ and Ⅱ was tested by SDS-PAGE and Western blot identification of Tembusu virus E protein domains I and Ⅱ expressed in Escherichia coli and induced by IPTG.The results showed that only a single target band was detected after purification.The titers of antibody of immunized group reached 4.67±0.11(lg10).The weight of immunized group was higher than that of control group from the 9th day postinoculation.Real-time RT-PCR showed that the Tembusu virus nucleic acid in the brain, liver and spleen of mice in immunized group was less than that of control group.It was indicated that the immunization with recombinant tembusu virus E protein domains I and Ⅱ could induce high level of specific antibody and provide immuno protection against Tembusu virus infection.
出处
《江苏农业学报》
CSCD
北大核心
2017年第2期-,共5页
Jiangsu Journal of Agricultural Sciences
基金
江苏省自然科学基金项目(BK20160064)
江苏省农业科技自主创新基金项目[CX(14)2091]
