不同启动子在昆虫杆状病毒表达系统中的活性分析  

Promoter Activity of Different Promoters in Recombinant Baculovirus-infected Sf9 Cells

作　　者：王煜[1] 高辉[1] 陈川[1] 赵和平[1] 李淼[1] 孙元[1] 仇华吉[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室猪传染病研究室

出　　处：《生物工程学报》2008年第4期598-603,共6页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No．2005CB523202)资助~~

摘　　要：为了比较不同启动子在杆状病毒/昆虫细胞中的活性,利用对虾白斑综合症病毒(White spot syndrome virus,wssv)的ie1启动子及其截短的mie1启动子、杆状病毒ETL启动子及其加长的mETL启动子以及杆状病毒多角体启动子P_(PH),构建了含有不同启动子控制下的EGFP报告基因的重组杆状病毒,分别感染Sf9昆虫细胞,利用流式细胞术检测报告基因的表达水平。结果表明,WSSV的ie1启动子和杆状病毒的mETL启动子在昆虫细胞sf9细胞中都具有较强而旱的启动子活性,能够控制报告基因早期高效表达,而P_(PH)在感染后期才表现较强活性。并且研究中发现,杆状病毒同源重复区(hr1)可以增强ETL启动子的活性。To compare the activity of different promoter in baculovirus-insect system, a series of recombinant baculoviruses were generated harboring the E-GFP reporter gene under the control of one of 5 promoters, including the iel promoter of shrimp white spot syndrome virus （WSSV）, the truncated iel （miel） promoter, the ETL promoter of the baculovirus, the elongated ETL （mETL） promoter, and the polyhedron promoter （Pen） of the baculovirus. The expression efficiency of the E-GFP reporter gene in the recombinant baculovirus-infected Sf9 cells was determined by flow cytometry. The results showed that both iel and mETL promoters had a strong promoter activity at early phase, while PPH showed a strong promoter activity at late phase. The iel promoter suggested the strongest promoter activity. The homologous region 1 （hr1） was also found to enhance the ETL promoter activity.

关 键 词：重组杆状病毒 启动子活性 流式细胞术 

分 类 号：Q78[生物学—分子生物学]