安徽农业科学2012，Vol.40Issue(16)：8932-8934,3.
牛轮状病毒结构蛋白VP4基因的真核表达
Eukaryotic Expression of Bovine Rotavirus Structural Protein VP4 Gene
摘要
Abstract
[ Qbjective ] To achieve the successful cloning and eukaryotic expression of the full-length VP4 gene of bovine rotavirus. [ Method ] VP4 gene was amplified from the total RNA in bovine rotavirus by RT-PCR technique, then cloned into pcDNA3. 1 ( + ) vector with Flag TAQ. The reeombinant vector was confirmed by PCR, restricting enzyme digestion and DNA sequence. The recorabinant plasmid was trans-fecled into 293T cells through Lipofect2000? The expressions of VP4 gene were confirmed by RT-PCR technique and Western Blot. [ Result] The reeombinant plasmid pcr)NA3. l( + )-VP4 was constructed, the detection by RT-PCR and Western Blot indicated thai the reeombinant plasmid pcDNA3.1( + )-VP4 has been constructed successfully and could express in 293T cells. [Conclusion] The reeombinant vector containing VP4 gene was constructed successfully and could express in eucaryotic cells. The study lays foundation for the development and application of DNA vaccine of VP4 gene.关键词
牛轮状病毒/VP4/基因克隆/真核表达Key words
Bovine rotavirus/ VP4/ Gene cloning/ Eukaryotic expression分类
农业科技引用本文复制引用
王丹,宋玲玲,王洪梅,杨宏军,王立群,何洪彬..牛轮状病毒结构蛋白VP4基因的真核表达[J].安徽农业科学,2012,40(16):8932-8934,3.基金项目
国家自然科学基金项目(31072160) （31072160）
国家转基因重大专项(2009ZX08007-006B) （2009ZX08007-006B）
济南市高校院所自主创新计划项目(201004027,201102034) （201004027,201102034）
BRV强毒株的致弱研究项目(Y2009-D56). （Y2009-D56）
