中国水产科学2016，Vol.23Issue(2)：352-358,7.DOI:10.3724/SP.J.1118.2016.15240
鲤春病毒血症病毒糖蛋白的截短表达及其免疫原性分析
Expression and immunogenicity analysis of a truncated glycoprotein of spring viremia of carp virus (SVCV)
摘要
Abstract
To reveal the main immunogenic domain of the glycoprotein of spring viremia of carp virus (SVCV), the glycoprotein encoding gene was truncated and expressed, and the immunogenicity of the truncated glycoprotein was analyzed using a rabbit polyclonal antibody produced in response to the recombination protein. The transmembrane, antigenic and hydrophilic domains of SVCV were analyzed by SOSUI and DNAStar 6.0 software. The predicted main immunogenic domain was amplified by reverse transcription polymerase chain reaction and cloned into a prokaryotic expression plasmid. The truncated glycoprotein was expressed in BL21 cells. After purification and refolding, the pro-tein was used to produce antiserum in rabbits. The antiserum titer was tested by an enzyme-linked immunosorbent assay and the immunogenicity of the protein was tested by immunoblotting and indirect immunofluorescence assay (IFA). The results showed that the truncated glycoprotein gene was 1339 bp encoding a 439 aa peptide (from 29 to 467aa of the full-length glycoprotein) with a predicted molecular weight of 49.6 kD. The antiserum titer against the recombinant glycoprotein was 1 : 64000. Immunoblotting and IFA results demonstrated that the antiserum reacted immunologically with the natural glycoprotein on the surface of SVCV-HN. There was no difference in immunogenicity between the truncated glycoprotein produced in this study and the natural glycoprotein of SVCV. Therefore, the truncated glycopro-tein of SVCV had good immunogenicity and could be used in immunological diagnosis and genetic engineering vaccine development of SVCV.关键词
鲤春病毒血症病毒/糖蛋白编码基因/克隆/蛋白截短表达/免疫原性Key words
spring viremia of carp virus (SVCV)/glycoprotein encoding gene/cloning/truncated expression/immu-nogenicity分类
农业科技引用本文复制引用
徐进,周勇,陈倩,曾令兵..鲤春病毒血症病毒糖蛋白的截短表达及其免疫原性分析[J].中国水产科学,2016,23(2):352-358,7.基金项目
现代农业产业技术体系建设专项资金资助项目(CARS-46-11) （CARS-46-11）
