福建农业学报Issue(5)：425-429,5.
鸭1型甲肝病毒亚型VP 3基因的克隆与原核表达
Cloning and Prokaryotic Expression ofVP 3 Gene of Duck Hepatitis A Virus Type 1 Subtype
摘要
Abstract
The VP 3 gene of the hepatitis A virus type 1 subtype (DHAV-1a)in ducks was amplified by the reverse transcription-polymerase chain reaction (RT-PCR)using one pair of specific primers designed according to the published sequences of DHAV-1a.The target DNA was purified and cloned into pEASYTM-Blunt Zero Cloning Vector.The recombinant expression plasmid,pET-32a-VP3,was constructed by inserting the target gene fragment into pET-32a (+)vector and transformed into Escherichia coli BL21 (DE3)competent cells.In this study,SDS-PAGE and Western-blot analyses showed that the recombinant protein VP 3,approximately 47 kDa in molecular mass,was expressed highly in E.coli after pET-32a-VP3 was induced with 1.0 mmol·L-1 IPTG at 37℃.The expressed recombinant protein was recognized specifically by Anti-His Mouse mAb showing a good bioactivity.关键词
鸭1型甲肝病毒亚型/VP3基因/克隆/原核表达Key words
duck hepatitis A virus type 1a/VP 3 gene/cloning/prokaryotic expression分类
农业科技引用本文复制引用
黄剑梅,黄瑜,傅秋玲,傅光华,万春和,陈翠腾,陈珍,程龙飞,施少华,陈红梅..鸭1型甲肝病毒亚型VP 3基因的克隆与原核表达[J].福建农业学报,2015,(5):425-429,5.基金项目
国家自然科学基金项目（31472222）；福建省科技计划项目---省属公益类科研院所基本科研专项（2014R1023-3）；现代农业产业技术体系建设专项（CARS-43）；福建省自然科学基金项目（2015J01113）；新世纪“百千万人才工程”国家级人选科研补助资金 （）
