摘要
以PPVVP2为基础,定点突变VP2蛋白中的Cys-402(A)、214(B),并构建突变体的真核表达载体,通过电镜技术和小鼠免疫试验探讨突变对VLPs的包装和免疫原性的影响。结果显示,在目标位点成功的引入突变,并构建了真核表达载体pPI-2-EGFP-VP2(A、B);电镜和小鼠免疫试验结果显示Cys-214对VLPs的组装和免疫原性的发挥是必需的,而Cys-402的突变并不干扰VLPs的形成和免疫原性的发挥,推测Cys-402及其附近可能存在利于外源基因插入的位点。本试验为以PPVVLPs为基础的多价疫苗和抗原转运系统提供了理论依据。
Based on the porcine parvovirus（PPV） VP2,site-directed mutation was imported on Cys-402（A） and Cys-214（B）,and then eukaryotic expression vectors of the mutants was constructed.Results：the mutations have been successfully imported and the eukaryotic expression vectors pPI-2-EGFP-VP2（A） and pPI-2-EGFP-VP2（B） were constructed;the result of electron microscope and BALB/c mice immuity test displayed that Cys-214 played an important role in VLPs＇ formation and immunogenicity;inversely,the mutation of Cys-402 was not to produce a marked effect on VLPs＇ formation and immunogenicity.It was supposed that there was a site which was of benefit to exogenous gene insertion on Cys-402 and nearby.This research provides theory reference to multivalent vaccine and antigen transport systems which are based on PPV VLPs.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2010年第3期294-298,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30500019)
四川省青年科技基金资助项目
四川省教育厅重点实验室专项
关键词
猪细小病毒
VP2
病毒样颗粒
定点突变
porcine parvovirus
VP2
virus like-particles
site-directed mutation
