| 山羊痘病毒双向启动子序列的克隆与鉴定 |
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| 引用本文： | 张娜,张辉,胡圣伟,王安涛,张沾,陈创夫,乔军.山羊痘病毒双向启动子序列的克隆与鉴定[J].中国预防兽医学报,2011,33(8). |
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| 作者姓名： | 张娜 张辉 胡圣伟 王安涛 张沾 陈创夫 乔军 |
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| 作者单位： | 石河子大学生命科学学院动物科技学院,新疆石河子,832000 |
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| 基金项目： | 国家973计划(2010CB30203); 国家自然科学基金(30800813) |
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| 摘 要： | 为鉴定山羊痘病毒(GPV)基因组中的启动子序列,本研究预测GPV基因组中早期转录因子VETF-l和中期转录因子VITF-3基因序列之间56 bp的一段序列为双向启动子,并将其命名为Pb56。通过PCR技术将该启动子序列的两个方向(Pb56+,Pb56-)分别与绿色荧光蛋白(EGFP)报告基因融合,构建重组转移重组质粒,分别命名为pUC-TK12-Pb56(+)-EGFP和pUC-TK12-Pb56(-)-EGFP。用脂质体转染法将2个转移重组质粒以及阴性对照pUC-TK12-EGFP及阳性对照pUC-TK12-P7.5-EGFP分别转染GPV预感染的BHK细胞;采用EGFP报告基因的表达水平评估双向启动子的活性。结果表明该基因序列的两个方向均能启动EGFP的表达,初步证实了该基因序列为GPV的双向启动子;Pb56+和Pb56-的转录活性均高于痘苗病毒的P7.5启动子。
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| 关 键 词： | 山羊痘病毒(GPV) 双向启动子 报告基因 P56b Pb56 转录活性 |
Identification of a bidirectional promoter from Goatpox virus |
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| ZHANG Na,ZHANG Hui,HU Sheng-wei,WANG An-tao,ZHANG Zhan,CHEN Chuang-fu,QIAO Jun.Identification of a bidirectional promoter from Goatpox virus[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(8). |
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| Authors: | ZHANG Na ZHANG Hui HU Sheng-wei WANG An-tao ZHANG Zhan CHEN Chuang-fu QIAO Jun |
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| Institution: | ZHANG Na,ZHANG Hui,HU Sheng-wei,WANG An-tao,ZHANG Zhan,CHEN Chuang-fu*,QIAO Jun*(College of Life Science,College of Animal Science and Technology,Shihezi University,Shihezi 832000,China) |
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| Abstract: | In this study,a 56 bp sequence between ETF-l and VITF-3 genes in Goatpox virus(GPV) genome was predicted to be a bidirectional promoter,and the sequence was synthesized and verified for bidirection of promoting activites by insert the GFP gene under control on either side of the 56 bp sequence,respectively.The recombinant transfer plasmids of pUC-TK12-Pb56(+)-EGFP and pUC-TK12-Pb56(-)-EGFP were constructed based on GPV TK gene as homologous arms and transfected into BHK cells preinfected with GPV,respective... |
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| Keywords: | Goatpox virus bidirectional promoter reporter gene promoter activity |
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