中国兽医科学2013，Vol.43Issue(3)：304-309,6.
脑心肌炎病毒VP1基因真核表达载体的构建及表达
Construction of eukaryotic expression vector system for expression of VP1 gene of encephalomyocarditis virus
摘要
Abstract
The objective of this study was to construct a eukaryotic expression vector harboring VP1 gene of encephalomyocarditis virus(EMCV) ,and to express the VP1 gene in CHO cells. VP1 gene segment of EMCV was amplified by RT-PCR,and then cloned into the pEGFP-Cl vector to construct eukaryotic expression vector pEGFP-C1-VP1. The constructed plasmid was then transfected into the CHO cell via Lipo-fectamineTM 2000 Reagent. Transcription of VP1 gene in the CHO cell was tested via RT-PCR,and localization and reactinogenicity of VP1 protein in the CHO cell were detected by using immunohistochemistry and Western-blot,respectively. Enzyme digestion of PCR products and sequencing showed that the recombinant plasmid pEGFP-Cl-VPl was successfully constructed. RT-PCR result showed that the VP1 gene was tran-scripted in the CHO cell. Immunohistochemistry result showed that the expressed VP1 protein was mainly distributed in the membrane of CHO cells, and a little in cytoplasm. Western-blot showed that the expressed VP1 protein could bind to rabbit anti-EMCV antibody,indicating the immunoreactivity of recombinant VP1 protein.关键词
脑心肌炎病毒/VP1基因/真核表达/免疫组织化学Key words
encephalomyocarditis virus/VP1 gene/eukaryotic expression immunohistochemistry分类
农业科技引用本文复制引用
凡静静,冯若飞,杨妍梅,张海霞,李向茸,王丹,谢晶莹,马忠仁..脑心肌炎病毒VP1基因真核表达载体的构建及表达[J].中国兽医科学,2013,43(3):304-309,6.基金项目
国家自然科学基金项目(31160033) （31160033）
中央高校基本科研业务费专项资金项目(ZYZ2012087) （ZYZ2012087）
西北民族大学科研创新项目中央专项(ycx12023) （ycx12023）
