【摘要】 目的:制备A和B亚群禽白血病病毒(Avian leukosis virus,ALV)特异性单克隆抗体。方法:用ALV-A-SDAU09C1株的env-gp85基因的PCR产物构建重组表达性质粒pET32a-SDAU09C1-gp85,经IPTG诱导后,表达分子量为53 kD的ALV-A囊膜gp85蛋白与GST的融合蛋白,将表达产物免疫BALB/c小鼠,取其脾脏细胞与骨髓瘤细胞SP2/0进行融合,筛选杂交瘤细胞株。结果:获得了1株(A6D1株)能与A和B亚群ALV发生反应但不与J亚群ALV反应的杂交瘤细胞株。Western blot试验结果表明,单克隆抗体识别的A和B亚群ALV囊膜糖蛋白的分子量为53 kD。在IFA中,这株单克隆抗体可以与所试验的3株ALV-A和1株ALV-B毒株反应,而与4株ALV-J亚群的毒株不反应。结论:A6D1株单克隆抗体可以用于A和B亚群ALV感染的诊断和流行病学调查,弥补了目前只有J亚群ALV特异性单克隆抗体可用的不足。更多还原

【Abstract】 Objective:Development and characterization of monoclonal antibody to subgroup A and B avian leukosis virus.Methods:The env-gp85 gene of ALV-A-SDAU09C1 was subcloned into the prokaryotic expressing vector pET32a and recombinant vector pET32a-SDAU09C1-gp85 was further transformed into Escherichia coli strain BL21 for expression under the induction of IPTG.After IPTG induction,there was a new protein band about 53 kD on SDS-PAGE.The expressed proteins were vaccinated into Balb/c mice.Results:One Mab(A6D1),which reacted with all exogenous subgroups of AL-AB but not with endogenous subgroup J,was obtained.Using immunofluorescence assay(IFA),A6D1 reacted with 3 ALV-A and 1 ALV-B,but not with 4 ALV-J.Western Blot showed that molecular weight of ALV AB envelope glycoprotein or unglycosylated envelope protein recognized by Mabs was about 53 kD.Conclusion:A6D1 can be used for diagnosis and epidemology of ALV-AB,which compensated for only the subgroup J ALV could be used in clinical.更多还原