中国动物传染病学报Issue(1)：1-8,8.
密码子优化型鸭甲肝病毒VP1基因在昆虫细胞中的表达
ENHANCED EXPRESSION OF CODON-OPTIMIZED VP1 GENE OF DUCK HEPATITIS A VIRUS IN INSECT CELLS
摘要
Abstract
The objective of the present study was to enhance expression level of VP1 gene of Duck hepatitis A virus (DHAV) genotype A in insect cells by manipulating the codon usage bias. The codon usage of wild-type DHAV VP1 (wtVP1) gene was optimized and designated as optiVP1. The recombinant rBacmid-optiVP1 and rBacmid-wtVP1 plasmids were then constructed using the Bac-to-Bac baculovirus expression system (BEVS) and then transfected to sf9 insect cells at logarithmic phase for expression of VP1 protein. The typical cytopathic effect was observed in sf9 cells at 72 h post transfection. The expression of VP1 protein in sf9 cells was confirmed in Western blotting and indirect immunofluorescence assay (IFA). The VP1 amounts on Western blotting were measured using the software Image J. The expression level of optiVP1 gene was significantly increased as compared with wtVP1 gene. This study provided a basis for development of diagnostic reagents and genetically engineered novel vaccines for DHAV.关键词
鸭甲肝病毒/VP1/重组杆状病毒/密码子优化Key words
Duck hepatitis A virus/VP1/recombinant baculovirus/codon optimization分类
农业科技引用本文复制引用
李传峰,陈宗艳,孟春春,梁瑞英,胡文,刘光清..密码子优化型鸭甲肝病毒VP1基因在昆虫细胞中的表达[J].中国动物传染病学报,2014,(1):1-8,8.基金项目
国家自然科学基金项目（31101848）；中央级公益性科研院所基本科研业务费专项（2012JB13）；公益性行业（农业）科研专项（201003012）；国家高技术研究发展计划（863）项目 （）
