猫杯状病毒VP1衣壳蛋白的截短表达及其抗原性分析  

Prokaryotic expression and antigenicity analysis of feline calicivirus truncated capsid protein VP1

作　　者：蒋艳妹 刘家森[2] 刘永相[2] 倪宏波[1] 曲连东[2] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国兽医科学》2016年第1期74-78,共5页Chinese Veterinary Science

基　　金：国家科技支撑计划项目(2013BAK11B01)

摘　　要：为筛选猫杯状病毒(FCV)VP1衣壳蛋白特异性优势抗原片段,利用生物信息学软件DNAStar对毒株2280的VP1衣壳蛋白各分区的抗原性进行了预测。在此基础上,以FCV 2280株cDNA为模板,RTPCR扩增VP1基因的A、B、CD、E和F共5个区段,构建重组表达质粒,5个基因片段在大肠杆菌中均获得高效可溶性表达。Western-blot试验证明,截短蛋白B、E、F均可与FCV阳性质控血清发生特异性反应,截短蛋白A和CD的信号弱。截短蛋白A、B、CD、E、F等量包被ELISA板,其中截短蛋白F与FCV阳性质控血清反应的D450,S/D450,N值最高,与其他蛋白之间差异显著(P<0.05)。结果表明,截短蛋白F为FCV VP1优势抗原片段,具备开发成FCV的血清学诊断试剂和新型疫苗的潜力。Bioinformatics software DNAStar was used to analyze the antigenicity of VP1 protein of feline calicivirus（FCV）strain 2280.And the gene sequences encoding fragments A,B,CD,E and F of VP1 protein were amplified by RT-PCR and cloned into vector pET-32a（＋）to construct the recombinant plasmids,respectively.The five target proteins were expressed as soluble proteins in Escherichia coli BL21（DE3）.Western-blot and ELISA were used to analysis the antigenicity of the purified protein.Proteins B,E and F were well-recognized the positive serum against FCV in Western-blot test,but proteins A and CD did not.In ELISA test,the D450,S/D450,Nvalue of the positive serum against protein F was the highest.In conclusion,the fragment F was proved to have the most antigen sites of FCV VP1 capsid protein,which could be applied to develop the serodiagnostic reagents and new vaccines against FCV.

关 键 词：猫杯状病毒 VP1蛋白 原核表达 抗原性 

分 类 号：S852.659.6[农业科学—基础兽医学]