【摘要】 为获得血清11型禽腺病毒(fowl adenovirus serotype 11,FAdV-11)纤突(Fiber)蛋白的多克隆抗体，并研究其对不同血清型FAdV Fiber蛋白的交叉反应特性，本研究利用同源重组技术将FAdV-11 Fiber蛋白的编码基因克隆至原核表达载体pET-32a上，转化至BL21 (DE3)感受态细胞，经诱导表达后，将纯化的重组蛋白作为免疫原免疫家兔制备多克隆抗体，并通过Western blot和IFA鉴定该多克隆抗体与不同血清型FAdV Fiber蛋白的交叉反应性。结果表明，His-FAdV-11-Fiber重组蛋白主要以包涵体形式表达且表达良好，Western blot和IFA结果均显示，制备的多克隆抗体能与FAdV-8a、FAdV-8b和FAdV-11的Fiber蛋白反应，而不能与FAdV-4的2个Fiber (Fiber 1和Fiber 2)蛋白反应。综上所述，本研究成功制备兔抗FAdV-11 Fiber的多克隆抗体，并验证其能特异性识别FAdV-8a、FAdV-8b和FAdV-11的Fiber蛋白，为进一步建立FAdV-11的血清学鉴别诊断方法奠定了基础。更多还原

【Abstract】 In order to obtain polyclonal antibodies against the fibrillar(Fiber) protein of fowl adenovirus serotype 11(FAdV-11) and investigate its cross-reactivity to different serotypes of FAdV Fiber, the gene encoding the FAdV-11 Fiber protein was cloned into a prokaryotic expression vector pET-32a by homologous recombination technology, then the plasmid was transformed into BL21(DE3) receptor cells, and the purified recombinant protein was used as an immunogen to immunize rabbits to prepare polyclonal antibody after induced expression, and the cross-reactivity of the polyclonal antibody against different serotypes of FAdV Fiber proteins was identified by Western blot and indirect immunofluorescence(IFA).The results showed that the His-FAdV-11-Fiber recombinant protein was mainly expressed as inclusion bodies and was well expressed.Western blot and IFA showed that the prepared polyclonal antibody reacted with the Fiber proteins of FAdV-8a, FAdV-8b, and FAdV-11,but did not with the 2 Fiber of FAdV-4(Fiber 1 and Fiber 2) proteins.In conclusion, in this study, we successfully prepared rabbit polyclonal antibodies against FAdV-11 Fiber and showed that it specifically recognized the Fiber proteins of FAdV-8a, FAdV-8b and FAdV-11,which lays the foundation for further establishment of serological differential diagnosis of FAdV-11.更多还原