| 虹鳟免疫诱导型基因Cathelicidin2启动子功能分析 |
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| 作者姓名： | 赵紫霞 许建 江炎亮 白庆利 蒋立坤 陈葆华 徐鹏 |
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| 作者单位： | 中国水产科学研究院农业农村部水生动物基因组学重点实验室渔业生物技术北京市重点实验室;中国水产科学研究院黑龙江水产研究所;厦门大学海洋与地球学院 |
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| 摘 要： | 本研究通过实时荧光定量PCR实验对虹鳟(Oncorhynchus mykiss)Cathelicidin2(Cath2)基因的转录模式进行了分析。结果显示,该基因在鳃、头肾等与机体免疫防御功能密切相关的组织内转录,在细菌和病毒感染后,转录水平均显著升高。对基因上游调控序列进行启动子和转录因子结合位点预测,发现该启动子具有真核生物典型的TATA盒和CAAT盒结构,基因上游直至第一内含子区域内,密集存在多个免疫相关转录因子结合位点,其中,2个核因子κB(Nuclear factor kappa B,NFκB)预测结合位点均位于核心启动子正链区域。在草鱼(Ctenopharyngodon idellus)肾组织细胞系内,绿色荧光蛋白和萤火虫荧光素酶基因都能够在该启动子驱动下表达,表明其具有启动子活性,且启动子活性在受到免疫诱导后增强,包括细菌脂多糖(Lipopolysaccharides,LPS)模拟的细菌感染和聚肌胞苷酸(Polyinosinic polycytidylic acid,Poly I:C)模拟的病毒感染。双荧光素酶报告基因检测显示,启动子活性在与NFκB转录因子表达时,增强至4.39倍,证明Cath2基因受NFκB通路调控。研究表明,虹鳟Cath2基因能够在多种免疫刺激诱导下表达,其启动子可以应用为免疫诱导型的基因工程元件,驱动外源免疫基因在鱼体内适时表达,抵御外界病原感染,同时,避免非必要条件下的过度表达形成生长负担。
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| 关 键 词： | 虹鳟 抗菌肽 Cathelicidin2 启动子 转录调控 |
| 收稿时间： | 2017/6/14 0:00:00 |
| 修稿时间： | 2017/7/7 0:00:00 |
Functional analysis of the immune-induced promoter of the rainbow trout Cathelicidin2 gene |
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| Authors: | ZHAO Zixi XU Jian JIANG Yanliang BAI Qingli JIANG Likun CHEN Baohua and XU Peng |
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| Institution: | Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Beijing Key
Laboratory of Fishery Biotechnology, Chinese Academy of Fishery Sciences, Beijing 100141,Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Beijing Key
Laboratory of Fishery Biotechnology, Chinese Academy of Fishery Sciences, Beijing 100141,Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Beijing Key
Laboratory of Fishery Biotechnology, Chinese Academy of Fishery Sciences, Beijing 100141,Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070,Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Beijing Key
Laboratory of Fishery Biotechnology, Chinese Academy of Fishery Sciences, Beijing 100141,Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Beijing Key
Laboratory of Fishery Biotechnology, Chinese Academy of Fishery Sciences, Beijing 100141 and Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Beijing Key
Laboratory of Fishery Biotechnology, Chinese Academy of Fishery Sciences, Beijing 100141; College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102 |
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