| 海带hsp70基因的克隆、分析及转录水平定量研究 |
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| 作者姓名： | 袁艳敏 刘福利 梁洲瑞 汪文俊 孙修涛 王飞久 |
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| 作者单位： | 上海海洋大学水产与生命学院;农业农村部海洋渔业可持续发展重点实验室中国水产科学研究院黄海水产研究所;青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 |
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| 摘 要： | 本研究以海带(Saccharina japonica)‘海天1号’为材料,通过RACE-PCR方法获得海带hsp70(Sjhsp70)基因全长cDNA序列。序列分析表明,全长cDNA长度为3778 bp,含有1个2892 bp的开放阅读框,5'和3'非编码区的长度分别为101、785 bp。蛋白质氨基酸含量和理化性质分析显示,该蛋白为稳定蛋白。应用BLASTP程序对不同物种HSP70蛋白比较分析表明,海带与长囊水云(Ectocarpus siliculosus)的HSP70蛋白具有较高的同源性,用HSP70蛋白构建的进化树显示,海带与长囊水云聚为一类。二级结构预测该蛋白由963个氨基酸组成,包含35个α螺旋和25个β折叠。以酿酒酵母(Saccharomyces cerevisiae)HSP110蛋白三维结构3C7N为模板,在I-TASSER软件上预测海带HSP70蛋白的三维结构,预测的蛋白结构与3C7N的A链在折叠和二级结构方面非常类似。对Sjhsp70基因在温度胁迫下表达变化分析表明,高温对Sjhsp70的表达量具有显著影响,Sjhsp70在25℃、24 h胁迫条件时,表达量达到最大值。本研究为阐明海带HSP70蛋白特性及其应对环境胁迫的机理提供理论依据。
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| 关 键 词： | 海带‘海天1号’ HSP70 RACE 生物信息学分析 荧光定量PCR |
| 收稿时间： | 2017/4/7 0:00:00 |
| 修稿时间： | 2017/5/8 0:00:00 |
Cloning, bioinformatics and quantitative expression analysis of hsp70 in Saccharina japonica |
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| Authors: | YUAN Yanmin LIU Fuli LIANG Zhourui WANG Wenjun SUN Xiutao and WANG Feijiu |
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| Institution: | College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306; Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese
Academy of Fishery Sciences, Qingdao 266071,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese
Academy of Fishery Sciences, Qingdao 266071; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266071,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese
Academy of Fishery Sciences, Qingdao 266071,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese
Academy of Fishery Sciences, Qingdao 266071,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese
Academy of Fishery Sciences, Qingdao 266071 and Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese
Academy of Fishery Sciences, Qingdao 266071 |
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