Construction and Application of Eukaryotic Expression Vector phIE Containing HSVI α-gene Promoter

CUN Wei; LIU Long-ding; ZHANG Ying; LUO Jie; WU Wen-juan; ZHANG Xue-mei; LIAO Yun; LI Qi-han*

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August 2006

CUN Wei, LIU Long-ding, ZHANG Ying, LUO Jie, WU Wen-juan, ZHANG Xue-mei, LIAO Yun and LI Qi-han*. Construction and Application of Eukaryotic Expression Vector phIE Containing HSVI α-gene Promoter[J]. Virologica Sinica, 2006, 21(4): 328-323.

Citation: CUN Wei, LIU Long-ding, ZHANG Ying, LUO Jie, WU Wen-juan, ZHANG Xue-mei, LIAO Yun, LI Qi-han*. Construction and Application of Eukaryotic Expression Vector phIE Containing HSVI α-gene Promoter .VIROLOGICA SINICA, 2006, 21(4) : 328-323.

利用HSV1 α-基因启动子构建真核表达载体phIE

1.
中国医学科学院中国协和医科大学医学生物学研究所，云南昆明 650118

摘要

以pcDNA3质粒为骨架，构建由1型单纯疱疹病毒(Herpes simplex virus I, HSVI)立即早期基因启动子指导外源蛋白转录的phIE真核表达载体，该质粒为外源蛋白的表达提供了更为广泛的选择，通过插入us1编码序列，phIE-Us1被用于分析ICP22蛋白在病毒感染早期的功能；在多克隆位点插入氯霉素乙酰转移酶编码基因构建phIE-CAT检测载体，应用于检测HSVI病毒蛋白Vp16及ICP22的功能，明确显示该质粒对于研究单纯疱疹病毒的基因转录调控具有重要的应用价值。
- 单纯疱疹病毒
- , 立即早期基因
- , 载体
- , 启动子

Construction and Application of Eukaryotic Expression Vector phIE Containing HSVI α-gene Promoter

1.
Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming 650118，China

Abstract

An eukaryotic expression vector phIE composed of pcDNA3 skeleton and the herpes simplex virus immediate early gene promoter was constructed to provide a new choice for ectopic expression. The plasmid phIE-Us1 was constructed by inserting the us1 cDNA downstream of the promoter to analyze the function of ICP22 protein very early in the infection process. Furthermore, the gene encoding the chloramphenicol acetyl-transferase was inserted into the multiple cloning site of the expression vector to yield the phIE-CAT. In functional analysis of viral protein ICP22 and Vp16, phIE-CAT was confirmed to be capable of evaluating the mechanism of the genome of herpes simplex virus 1 transcriptional regulation.
- Herpes simplex virus
- , Immediate early gene
- , Vector
- , Promoter

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction and Application of Eukaryotic Expression Vector phIE Containing HSVI α-gene Promoter

1. Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming 650118，China

Abstract: An eukaryotic expression vector phIE composed of pcDNA3 skeleton and the herpes simplex virus immediate early gene promoter was constructed to provide a new choice for ectopic expression. The plasmid phIE-Us1 was constructed by inserting the us1 cDNA downstream of the promoter to analyze the function of ICP22 protein very early in the infection process. Furthermore, the gene encoding the chloramphenicol acetyl-transferase was inserted into the multiple cloning site of the expression vector to yield the phIE-CAT. In functional analysis of viral protein ICP22 and Vp16, phIE-CAT was confirmed to be capable of evaluating the mechanism of the genome of herpes simplex virus 1 transcriptional regulation.