含有增强型绿色荧光蛋白报告基因的辛德毕斯病毒的制备与鉴定  

Construction and characterization of EGFP reporter gene labeled Sindbis virus

作　　者：邓玲玲[1] 李江姣[2] 尉研[2] 王焕琴[2] 张凤娟[2] 孙继国[2] 陈畅[1] 朱武洋[2] 梁国栋[2] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029 [2]中国疾病预防控制中心病毒病预防控制所脑炎室

出　　处：《中华实验和临床病毒学杂志》2013年第3期228-230,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的 制备含有增强型绿色荧光蛋白（enhanced green fluorescent protein,EGFP）报告基因的辛德毕斯病毒,并对其进行鉴定.方法 采用融合PCR技术将报告基因EGFP融合到辛德毕斯病毒XJ-160感染性克隆pBR-XJ160的结构基因编码框后,并在报告基因前面添加亚基因启动子SP6,通过反向遗传学技术拯救得到EGFP标记的辛德毕斯病毒.结果 成功拯救并获得了EGFP标记的XJ-160病毒,该重组病毒具有良好的荧光表达特性和遗传稳定性.结论 EGFP标记的辛德毕斯病毒可用于观测病毒的侵染轨迹,为进一步研究辛德毕斯病毒嗜性、生物学功能,以及感染机制奠定了基础.Objective To construct and characterize EGFP reporter gene labeled Sindbis virus （SINV）.Methods The reporter gene EGFP was inserted into the genome of infectious clone pBR-XJ160 by using multi-fusion long fragment PCR method.Then apply reverse genetic manipulation technique to rescue and obtain EGFP labeled SINV.Results We successively obtained labeled SINV,which has good fluorescent expression characteristics and genetic stability.Conclusion The labeled virus can be seen in living cells and living body,and this serves as a good tool for cell and tissue tropism and biological function study of viruses.This study laid a foundation for further studying the cell tropism,biological functions and infection mechanism of SINV.

关 键 词：Sindbis病毒 基因 报告 克隆 分子 

分 类 号：R[医药卫生]