摘要
为研究猪圆环病毒2型(PCV2)ORF2基因在异源细胞上的表达及生物学特性,构建了PCV2ORF2与增强型绿色荧光蛋白(EGFP)的融合基因真核表达载体pEGFP-N1-ORF2。采用脂质体法转染体外培养的鸭心肌细胞后,通过直接荧光观察EGFP-ORF2融合蛋白在鸭心肌细胞中的分布定位,经RT-PCR、间接免疫荧光方法检测ORF2基因在鸭心肌细胞中的表达。结果显示,转染48 h后有绿色荧光出现,RT-PCR和间接免疫荧光法检测均为阳性。证实,PCV2 ORF2基因在鸭心肌细胞内获得了成功表达。
In this study,ORF2 gene of PCV2 was cloned via PCR method,and the eukaryotic expression plasmid of ORF2 gene with enhanced green fluorescent protein（EGFP） report gene was constructed.The recombinant plasmid was transfected with lipofectin method prepared duck myocardial cells（DMCs）.The fluorescence expression was directly detected with fluorescence microscope,and the expression of ORF2 were tested by RT-PCR and indirect immunofluorescence assay（IFA）,respectively.About 48h after transfected,green fluorescent can be detected in transfected cells,and RT-PCR and IFA were positive.The results indicated that ORF2 gene was expressed efficiently in transfected duck myocardial cells.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2011年第10期1047-1051,共5页
Chinese Veterinary Science
基金
山东省自主创新成果转化重大专项计划项目(2008ZHZX1A1103)
