虎源FPV主要抗原表位区基因的原核表达及免疫原性分析  

Prokaryotic Expression of FPV Major Epitope Domain and Analysis of Its Immunogenicity

作　　者：杨莹莹[1] 田丽红[1] 曾祥伟[1] 

机构地区：[1]东北林业大学野生动物资源学院,哈尔滨150040

出　　处：《经济动物学报》2014年第1期24-28,共5页Journal of Economic Animal

基　　金：国家林业局野生动植物保护管理项目(2011)

摘　　要：为筛选出制备猫瘟热亚单位疫苗的最佳佐剂类型与首选抗原片段,本试验对虎源FPV-HLJ株VP2全长基因及其截短基因片段PAB进行原核表达,切胶法纯化的表达蛋白经Western blot分析后,分别与氢氧化铝胶佐剂及弗氏佐剂混合,免疫BALB/c小鼠,间接ELISA方法检测小鼠体液免疫水平,初步评价各疫苗的免疫效果。结果表明:VP2、PAB融合蛋白主要以包涵体形式表达,切胶法获得的高纯度表达蛋白均能与鼠抗虎源FPV阳性血清发生特异性反应。纯化蛋白各佐剂免疫组免疫小鼠后均引起较强的特异性抗体IgG反应,其中PAB蛋白各免疫佐剂组抗体水平明显高于VP2蛋白相应免疫佐剂组,且PAB蛋白+氢氧化铝胶组与PAB蛋白+弗氏佐剂组抗体水平无显著差异(P>0.05),但与未加佐剂组差异显著(P<0.01)。研究证实,PAB蛋白具有良好的免疫原性,可与氢氧化铝胶佐剂协同用于FPV亚单位疫苗的制备。To study the best adjuvant and antigen of subunit vaccines of Feline Panleukopenia virus, the gene fragnlents VP2, PAB of FPV-HIJ isolated from Siberian tigers were expressed in E. coli BL21 （DE3）. Western blot method was used to detect the antigenicity of the target proteins which were ob- tained by cutting the gel slices contained the right band. Then, the purified proteins were mixed with the AI（OH）3 adjuvant and Freund＇ s adjuvant to apply to immune the BALB/c mice. To evaluate the effect of the vaccines preliminarily, the antibody was detected by indirect ELISA. The results showed that the fu- sion protein VP2 and PAB were highly expressed in the form of inclusion body. They could be identified by the polyclonal antibody from the mouse against tiger Feline distemper virus. All the adjuvant groups were able to enhance specific IgG response. The immune effect of the purified PAB protein was superior to VP2. And antibody titers were no significant difference between PAB protein ＋ AI（OH）3 adjuvant group and PAB protein ＋ Freund＇ s adjuvant group （ P 〉 0.05）, but there was a significant difference compared with the no adjuvant group （P 〈 0.01 ）. The study confirmed that PAB protein had good im- munogenieity and could be synergistic with AI（OH）3 adjuvant for the FPV subunit vaccine.

关 键 词：猫瘟热病毒 原核表达 VP2蛋白 免疫原性 

分 类 号：S858.315.3[农业科学—临床兽医学]