中国人兽共患病学报2017，Vol.33Issue(1)：32-37,6.DOI:10.3969/j.issn.1002-2694.2017.01.006
4种血清型登革病毒多表位重组抗原的表达及血清学初步评价
Expression and its serological preliminary evaluation on multi-epitope recombinant antigens of 4 serotypes of dengue virus
易方浩 1郑碧英 1徐军发 1张俊爱 1黎四平 2贾岩 1陈晨 1余诗炎 1王鑫 1代友超 1庄泽岗1
作者信息
- 1. 广东医科大学检验医学研究所,东莞 523808
- 2. 东莞市第八人民医院,东莞 523808
- 折叠
摘要
Abstract
We expressed B cell epitopes of dengue virus envelope protein and NS1 protein in prokaryotic cells,and purified and evaluated for its serological activities.A recombinant multi-epitope chimeric gene named rE including eight B cell epitopes was connected by linker peptide (EAAAK)2 and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The expressed recombinant protein was purified with 6× His purification media,and identified by SDS-PAGE and Western blot,and its antigenicity was analyzed by using an indirect ELISA assay.The recombinant expression vector pET28a-rE was constructed and expressed in BL21 (DE3) successfully,but the recombinant proteins mainly appeared as inclusion bodies.The target protein was obtained with high purity through the purification of affinity chromatography.SDS-PAGE and Western blot analysis showed that the molecular weight of fusion protein was in the expected line.The established indirect ELISA has high accuracy.This recombinant peptide antigen expressed in E.coli has good potential for serum testing.关键词
登革病毒/抗原蛋白/原核表达/血清学评价Key words
dengue virus/antigen protein/prokaryotic expression/serological evaluation分类
医药卫生引用本文复制引用
易方浩,郑碧英,徐军发,张俊爱,黎四平,贾岩,陈晨,余诗炎,王鑫,代友超,庄泽岗..4种血清型登革病毒多表位重组抗原的表达及血清学初步评价[J].中国人兽共患病学报,2017,33(1):32-37,6.
