【摘要】 以重组质粒 p GEX-MDg I在大肠杆菌 BL2 1中表达马立克氏病病毒 (MDV)特超强毒 64 8A株的囊膜糖蛋白 I(g I)完整基因 ,通过 SDS-聚丙烯酰胺凝胶电泳分离相应的蛋白条带 ,切下并碾碎后作为免疫原免疫小鼠 ,用以制备针对MDV64 8A g I糖蛋白的杂交瘤细胞株。以 MDV感染的鸡胚成纤维细胞 (CEF)为检测抗原 ,用间接免疫荧光抗体反应(IFA)筛选到 9株杂交瘤细胞株 ,选其中 3株 2 H7、6F1 0、2 H3制备腹水 ,其 IFA效价均达到 1∶ 1 50 0以上 ;与 I型 MDV不同致病型的毒株 CVI988/ Rispens、GA、RB1 B、Md1 1株感染的 CEF测 IFA,均呈现特异性荧光 ,但反应强度不同。腹水经 PBS稀释后与 MDV感染的 CEF做斑点酶联免疫吸附试验 ,呈特异性显色。对 MDV感染的 CEF裂解物做 West-ern-blotting,可检测出 1条特异性条带更多还原

【Abstract】 The fusion product of glycoprotein I of very virulent plus (vv +) MDV 648A strain expressed in Escherichia coli was discerned and excised from SDS PAGE, then inoculated BALB/C mice directly as immunogen to prepare specific monoclonal antibody. 3 strains of monoclonal antibody hydridoma cells were obtained by IFA, and all were specific to MDV infected CEF through dot ELISA. This is the first report on gI Mab that is responsive to MDV in cell cultures through Western blotting. These Mabs might be used to elicit the possible relationships between MDV variation and expression of gI in virus infected cells.更多还原