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中文摘要：
为鉴定牛传染性鼻气管炎病毒（IBRV）gD蛋白抗原表位，本研究利用pET-30a原核表达系统表达、纯化了重组gD蛋白，采用超速离心纯化的IBRV免疫BALB／c小鼠，取脾淋巴细胞与SP2／0细胞进行融合，以重组gD蛋白作为包被抗原，通过间接ELISA筛选出一株稳定分泌抗gD蛋白的MAb杂交瘤细胞株（1G3）。间接免疫荧光结果表明：MAb与IBRV呈阳性反应，中和试验测定其腹水中和效价为1：32。利用肽扫描技术原核表达GST融合短肽对MAb1G3识别的抗原表位进行筛选鉴定，结果表明：该MAb识别的抗原表位为259 EESKGyE265。蛋白序列分析表明该抗原表位在IBRV各分离株及牛疱疹病毒5型（BoHV-5）中均保守。本研究为IBRV的糖蛋白gD的结构和免疫学特性及IBRV致病性的深入研究奠定了基础。
英文摘要：
To identify the epitope on glycoprotein D （gD） of infectious bovine rhinotracheitis virus （IBRV）, the SP2/0 cells were fused with spleen cells from BALB/c mice immunized with purified IBRV, and one hybridoma stably secreting monoclonal antibody （MAb） against gD of IBRV was identified by indirect ELISA coated with gD expressed in E.coli. The MAb was positively reacted with IBRV detected by indirect immunofluorescence assay. The titer of the MAb prepered in ascites was 1：32 by virus neutralization test. The antigen epitope was further identified with the MAb by the pepscan method using GST-fused peptides expressed in E.coli, and a linear epitope of 259EESKGYE265 was identified in the gD. Moreover, the deduced amino acid sequence alignment of IBRV gD sequences showed that the epitope was completely conserved among different isolates of IBRV and BoHV-5. The identification of the epitope would contribute to further study of the structure and immunogenicity of IBRV gD and the pathogenicity of IBRV.
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