中国兽医科学2018，Vol.48Issue(5)：574-579,6.DOI:10.16656/j.issn.1673-4696.2018.0076
H3N2亚型犬流感病毒单链抗体的制备
Preparation of single-chain antibody against H3N2 canine influenza virus
摘要
Abstract
To prepare the single-chain antibody (scFv) against H3N2 canine influenza virus (CIV), total RNA was extracted from the spleens of BALB/c mice immunized with purified CIV,and the gene fragments of heavy chain(VH) and light chain variable regions(VL) were amplified by PCR. The VH-Linker-VL fusion fragment was constructed by ligating the VH and VL through the elastic peptide Linker. The recombinant plasmid was constructed by inserting the target fragment into the expression vector pET-32a(+ ),and positive clones were transformed into Escherichia coli BL21 (DE3). After induced by IPTG,the target protein band with a size of about 45 ku was obtained. Western-blotting analysis showed that the protein could be specifically recognized by anti-His Tag mouse monoclonal antibody. Indirect ELISA and neutralization test results showed that the scFv could specifically react with CIV antigen and had neutralizing activity.In this study,the scFv against H3N2 CIV was successfully prepared,which provided the material basis for the diagnosis and treatment of CIV infection.关键词
犬流感病毒/单链抗体/原核表达/中和活性Key words
canine influenza virus/single-chain antibody/prokaryotic expression/neutralizing activity分类
农业科技引用本文复制引用
邱冬,赵丹,刘永杰..H3N2亚型犬流感病毒单链抗体的制备[J].中国兽医科学,2018,48(5):574-579,6.基金项目
国家重点研发计划项目(2017YFD0501101) （2017YFD0501101）
科技部国际合作专项(2014DFG32770) （2014DFG32770）
