利用RNA复制子融合表达口蹄疫病毒抗原表位与分子内佐剂的研究及其表达产物的免疫学分析  

Expression of foot-and-mouth disease virus epitopes fused with intramolecular adjuvant using RNA replicon and immunological analysis of the expressed product

作　　者：朱良隆 毛箬青[1] 王相伟 殷相平[1] 孙跃峰 ZHU Liang-long;MAO Ruo-qing;WANG Xiang-wei;YIN Xiang-ping;SUN Yue-feng(State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2021年第5期537-544,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2016YFE0204100)。

摘　　要：为了探究新型疫苗载体在口蹄疫防控中的应用,本研究构建了一种融合表达口蹄疫病毒抗原表位与分子内佐剂的RNA复制子。首先将FMDV毒株O/BY/2010 B、T表位及霍乱毒素B和α-干扰素基因构建到RNA复制子载体中,通过将复制子质粒线性化并进行体外转录,得到复制子RNA。将该RNA转染BHK-21细胞,用Western-blot以及RT-PCR验证其正确表达后,选择不同佐剂免疫小鼠,用ELISA检测特异性抗体的产生。结果,重组复制子能够在BHK-21细胞内正常表达,免疫小鼠后能够诱导产生特异性抗体。综上所述,本研究构建的重组RNA复制子能够表达FMDV抗原表位与分子内佐剂,为口蹄疫RNA疫苗的研制提供了理论基础。In order to study the application of new vaccine carriers in the prevention and control of foot-and-mouth disease,a self-replication RNA vaccine containing FMDV epitopes and intramolecular adjuvants was developed in this study.Firstly,the B and T epitopes of FMDV O/BY/2010 strain and the cholera B and interferon-α genes were constructed into the RNA replicon vector.The replicon RNA was obtained by in vitro transcription of linearizing the replicon plasmid.Secondly,the RNA was transfected into BHK-21 cells.After the expressions of epitopes and intramolecular adjuvants were verified by Western-blot and RT-PCR,optimal adjuvants were selected to prepare vaccine.Mice were immunized by the RNA vaccine,and the titer of specific antibody was quantified by ELISA assay.In result,the gene of interest in the recombinant replicon were expressed successfully in BHK-21 cells,and the specific antibody could be detected in serum of the immunized mice.In summary,the recombinant RNA replicons constructed in this study could express FMDV epitopes and intramolecular adjuvants,which provide a clue to develop a new FMD RNA vaccine.

关 键 词：RNA复制子 口蹄疫病毒 表位 分子内佐剂 

分 类 号：S852.659.6[农业科学—基础兽医学]