摘要
口蹄疫病毒接种乳鼠、豚鼠及BHK-21细胞,获得相应的适应毒。以各组织液中的口蹄疫病毒RNA为模板,反转录并扩增VP1基因,PCR产物琼脂糖凝胶电泳,回收目的片段,测序。测序结果与核酸数据库中口蹄疫病毒VP1基因比对,并将各适应毒扩增的VP1基因序列相互比对。结果表明:此毒为口蹄疫病毒,各适应毒间序列同源性达98%。并用口蹄疫病毒抗原进行SDS-PAGE,分别与大肠杆菌表达的口蹄疫病毒HeB株P1蛋白制备的多克隆兔抗及健康对照血清进行Western blot,对多克隆兔抗进行鉴定,并用间接ELISA测定此多克隆兔抗的效价。确定此多抗能中和FMDV结构蛋白,抗体效价>1∶800。
Foot and mouth disease virus （FMDV） Asia I/HeB strain was inoculated into mice, guinea-pigs and BHK- 21 cell line to obtain corresponding adapted virus. The RNA of FMDV in each tissue fluid was taken as template for reverse transcription and amplification of VP1 gene, and then the agarose gel electrophoresis was conducted to the PCR product to collect aimed segment for sequencing. The sequencing results were compared with FMDV VP1 gene in the GenBank, and the sequences of each adapted virus amplified VP1 gene were compared with each other. The results showed that this virus was identified as FMDV, and the homology of sequence among the three host-adapted viruses reached 98%. The antigen of FMDV was used to conduct SDS-PAGE that it was used to conduct Western blot with polyclonal antibody, which was prepared by inoculating E. coll. expressed FMDV HeB strain P1 protein into rabbit, and control （healthy serum）, respectively in order to identify polyclonal antibody from rabbit. Moreover, the indirect ELISA was used to determine titer of this rabbit polyclonal antibody. The results confirmed that this rabbit polyclonal antibody can neutralize FMDV structural protein and its titer is more than 1：800.
出处
《湖南农业科学》
2012年第4期126-129,共4页
Hunan Agricultural Sciences
基金
十二五"863"计划(2011AA10A211)
关键词
口蹄疫病毒
结构蛋白P1
多克隆抗体
foot and mouth disease virus （FMDV）
structural protein P1
polyclonal antibody
作者简介
作者简介：郭少娟（1984－），女，河南登封市人，硕士，研究方向为动物疫苗及分子免疫学。
通讯作者：张永光
