【摘要】 本研究旨在建立一种生产水貂阿留申病(Aleutian disease,AD)诊断抗原的新方法。试验提取水貂阿留申病毒(Aleutian disease virus,ADV)的基因组,PCR扩增ADV核衣壳蛋白VP2基因,构建重组表达质粒BacmidVP2,脂质体介导其转染昆虫细胞Sf9获得重组杆状病毒AcMVPV-VP2。电镜下观察表达的VP2蛋白,Western blotting检测目的蛋白的反应原性。以传统接毒方法生产的AD诊断抗原作对照,通过对流免疫电泳试验检测表达蛋白的生物学活性。结果表明,表达的重组VP2蛋白在电镜下组装成病毒样颗粒且能与ADV阳性血清发生反应。与商业诊断抗原相比,重组抗原诊断AD的阴阳性的符合率为100%。该方法可成为生产AD诊断抗原的替代方法。更多还原

【Abstract】 To develop a new method of Aleutian disease(AD)diagnostic antigen production,we used Bac-to-Bac expression system in this study.Firstly,Aleutian disease virus(ADV)genome was extracted and ADV VP2 gene was amplified by PCR method.Then Bacmid-VP2 was constructed and transfected into insect cell Sf9 by liposomes to construct AcMVPV-VP2.Secondly,the VP2 protein was observed by electromicroscope and antigency was detected by Western blotting.At last,the activity of recombinant protein was inspected by countercurrent immunoelectrophoresis.The results showed that the expressed recombinant VP2 protein could react with ADV positive serum and form virus like particles.Compared with the commercial diagnostic antigen,the coincidence rate of recombinant antigen was 100%.This method could be a candidate for AD diagnostic antigen production.更多还原