Kumar S S, Renji R, Saini M, Goel A C, Sharma B
Division of Biochemistry and Food Science, Indian Veterinary Research Institute, Izatnagar, India.
Biochem Mol Biol Int. 1998 Dec;46(6):1093-100. doi: 10.1080/15216549800204642.
A cDNA library of Rinderpest vaccine virus was prepared in Zap Express vector (Stratagene). The Rinderpest 'N' gene specific clones were selected, characterized and thereafter expressed in E. coli XLOLR strain. The expressed protein was found to be immunogenic in western blot with hyperimmune sera. It reacted with rinderpest and 'N' protein specific monoclonal antibodies in Enzyme Linked Immunosorbent Assay (ELISA). Prokaryotically expressed 'N' protein also gave precipitin band in counter immunoelectrophoresis test (CIE). The expression of N protein was sufficient for its utility as positive antigen in CIE and ELISA used for rinderpest diagnosis.
用Zap Express载体（Stratagene公司）制备了牛瘟疫苗病毒的cDNA文库。筛选出牛瘟“N”基因特异性克隆，进行鉴定，随后在大肠杆菌XLOLR菌株中表达。经检测，表达的蛋白在用超免疫血清进行的蛋白质免疫印迹试验中具有免疫原性。在酶联免疫吸附测定（ELISA）中，它能与牛瘟和“N”蛋白特异性单克隆抗体发生反应。原核表达的“N”蛋白在对流免疫电泳试验（CIE）中也出现了沉淀带。N蛋白的表达足以使其作为用于牛瘟诊断的CIE和ELISA中的阳性抗原。
