| 不同启动子在昆虫杆状病毒表达系统中的活性分析 |
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| 引用本文： | 王煜,高辉,陈川,赵和平,李淼,孙元,仇华吉.不同启动子在昆虫杆状病毒表达系统中的活性分析[J].微生物学报,2008,24(4):598-603. |
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| 作者姓名： | 王煜 高辉 陈川 赵和平 李淼 孙元 仇华吉 |
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| 作者单位： | 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, 哈尔滨 150001; 内蒙古农业大学动物科学与医学学院, 呼和浩特 010018;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, 哈尔滨 150001;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, 哈尔滨 150001;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, 哈尔滨 150001;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, 哈尔滨 150001;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, 哈尔滨 150001;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, 哈尔滨 150001 |
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| 基金项目： | 国家自然科学基金(No. 2005CB523202) 资助。 |
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| 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PPH, 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早的启动子活性, 能够控制报告基因早期高效表达, 而PPH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。
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| 关 键 词： | 重组杆状病毒 启动子活性 流式细胞术 |
Promoter Activity of Different Promoters in Recombinant Baculovirus-infected Sf9 Cells |
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| Yu Wang,Hui Gao,Chuan Chen,Heping Zhao,Miao Li,Yuan Sun and Huaji Qiu.Promoter Activity of Different Promoters in Recombinant Baculovirus-infected Sf9 Cells[J].Acta Microbiologica Sinica,2008,24(4):598-603. |
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| Authors: | Yu Wang Hui Gao Chuan Chen Heping Zhao Miao Li Yuan Sun and Huaji Qiu |
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| Institution: | Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China; College of Animal Sciences and Veterinary Medicine, Inner Mon;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China |
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| Abstract: | 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 ie1 promoter of shrimp white spot syndrome virus (WSSV), the truncated ie1 (mie1) promoter, the ETL promoter of the baculovirus, the elongated ETL (mETL) promoter, and the polyhedron promoter (PPH) 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 ie1 and mETL promoters had a strong promoter activity at early phase, while PPH showed a strong promoter activity at late phase. The ie1 promoter suggested the strongest promoter activity. The homologous region 1 (hr1) was also found to enhance the ETL promoter activity. |
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| Keywords: | recombinant baculoviruses promoter activity flow cytometry |
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