中国兽医科学2018，Vol.48Issue(2)：175-181,7.DOI:10.16656/j.issn.1673-4696.2018.0027
伪狂犬病病毒gB抗原的可溶性原核表达及纯化条件的优化
Prokaryotic expression of pseudorabies virus gB gene and optimization of protein purification conditions
摘要
Abstract
In order to study the immunogenicity of pseudorabies virus gB protein,we constructed the prokaryotic expression system containing gB gene,and named as recombinant plasmid pSMK-gB.It was transformed into Escherichia coli and induced to express target protein.The induction temperature and time,protein solubility,and final concentration of IPTG were optimized.The results showed that the pseudorabies virus gB protein was expressed at the highest level,when temperature was 16 ℃,the D600 value was up to 1.2,IPTG concentration was 0.8mmol/L,and termination of induction at 20 h.In addition,the expressed protein was soluble.After purification of gB protein using nickel column,Western-blot analysis was used to confirm the target protein with anti-histidine tag specific antibody as primary antibody.This study will provide the condition for development of the gB-based ELISA kit in the future.关键词
伪狂犬病病毒/gB基因/可溶性表达/原核表达Key words
pseudorabies virus/gB gene/soluble expression/prokaryotic expression分类
农业科技引用本文复制引用
李娇阳,董虎,郭慧琛,孙世琪,张小丽..伪狂犬病病毒gB抗原的可溶性原核表达及纯化条件的优化[J].中国兽医科学,2018,48(2):175-181,7.基金项目
国家国际科技合作专项(2014DFA31890) （2014DFA31890）
国家自然科学基金项目(31672592) （31672592）
中央级科研院所基本科研业务费专项(1610312016002,1610312017010) （1610312016002,1610312017010）
中国农业科学院青年英才计划项目 （）
国家重点研发计划项目(2017-YFD0500906) （2017-YFD0500906）
