中国动物传染病学报2016，Vol.24Issue(1)：32-37,6.
猪流感病毒新型核糖体移码蛋白PA-X特有序列的原核表达、纯化及鉴定
PROKARYOTIC EXPRESSION, PURIFICATION AND IDENTIFICATION OF PA-X-SPECIFIC SEQUENCE OF SWINE INFLUENZA VIRUS
摘要
Abstract
The novel influenza A virus protein PA-X is a ribosomal frame-shifting product that modulates the host response and viral virulence. In the present study, PA-X protein was prokaryotically expressed, purified and characterized in order to investigate its biological functions. The PA-X coding gene was amplified in RT-PCR and overlapping PCR. The prokaryotic expression vectors were constructed with the gene recombination technique and verified using bacterium solution PCR and sequencing. The positive clones were transformed into E.coli BL21 (DE3) and the expressed protein was analyzed for its solubility. The PA-X products were purified using nickel column chromatography. The sequences of the plasmids and open reading frames were demonstrated to be completely correct. After transformation, these plasmids expressed the expected proteins with soluble nature. Subsequently, the PA-X protein was purified and used in Western blot to react with the mouse antiserum against the H1N1 swine influenza virus. The positive reaction between the PA-X protein and mouse antiserum was observed.关键词
猪流感病毒/PA-X特有序列/原核表达/蛋白纯化/Western blotKey words
Swine influenza virus/PA-X specific sequence/prokaryotic expression/purified protein/Western blot分类
农业科技引用本文复制引用
宫晓倩,郑浩,童光志,于海,阮宝阳,刘晓敏,张鹏,汪琪,汪秀会,周艳君,李泽君,童武..猪流感病毒新型核糖体移码蛋白PA-X特有序列的原核表达、纯化及鉴定[J].中国动物传染病学报,2016,24(1):32-37,6.基金项目
国家青年自然科学基金（31201916）；上海市自然科学基金青年项目（12ZR1453500）；中央级公益性科研院所基本科研业务费项目（2015JB07）；中国农业科学院创新工程科研团队“动物流感病毒病原生态学”项目 （）
