中国兽医科学2017，Vol.47Issue(6)：671-679,9.DOI:10.16656/j.issn.1673-4696.2017.06.001
HN蛋白携带TC标签的重组新城疫病毒的构建及鉴定
Construction and identification of recombinant Newcastle disease virus containing TC tagged HN protein
摘要
Abstract
Newcastle disease virus(NDV) is the primary cause of morbidity and mortality of poultry worldwide.As an important envelope glycoprotein,HN protein plays essential roles in the course of the adsorption and entry process of NDV.In this study,the TC(Tetracysteine) tag which is composed of six amino acids was introduced into the N-terminal of coding region of HN gene through reverse genetic technology.The TC tagged recombinant virus La Sota-HN-NTC was rescued in transfected cells,and this recombinant virus could effectively replicate in chicken eggs,of which the growth kinetics was similar to parental La Sota.The result of Western blot showed that there was no obvious difference of expression of HNprotein between La Sota-HN-NTC and La Sota.The virulence of La Sota HN-NTC was slightly lower than La Sota,and the TC tag could stably exist in recombinant virus.Green fluorescence was observed in La Sota-HN-NTC infected cells,suggesting that TC tagged HN protein could be expressed and specifically stained by biarsenical dye.The adsorption assay showed that the ability of La Sota-HN-NTC attachment to cellular receptor did not affected by TC tag.Furthermore,the dynamic distribution of HN in La Sota-HN-NTC infected cells was monitored by laser confocal microscopy.The present study provides the tool and basis for elucidating the dynamic distribution and the mechanism of HN during the infection process of NDV.关键词
新城疫病毒/TC标签/HN蛋白Key words
Newcastle disease virus/TC tag/HN protein分类
农业科技引用本文复制引用
齐甜铭,赵文钧,陈玉秋,孙军峰,马得莹..HN蛋白携带TC标签的重组新城疫病毒的构建及鉴定[J].中国兽医科学,2017,47(6):671-679,9.基金项目
黑龙江省应用技术研究与开发计划项目(PC13S02) （PC13S02）
国家自然科学基金项目(31502088) （31502088）
黑龙江省自然科学基金项目(QC2015038) （QC2015038）
