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| 魁蚶Ets家族9个基因的克隆及其在病毒感染应答中的表达分析 |
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魏智薪1,2, 辛鲁生3,2, 白昌明3,2, 李亚楠2, 张淑敏2, 李成华1, 王崇明3,2
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1.宁波大学海洋学院 宁波 315211;2.中国水产科学研究院黄海水产研究所 农业农村部海水养殖病害防治重点实验室 青岛市海水养殖流行病学与生物安保重点实验室 青岛 266071;3.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 青岛 266071
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| 摘要: |
| Ets蛋白是宿主MAPK信号通路下游的一类可参与调控病毒基因转录复制的重要转录因子。本研究通过基因克隆成功获得魁蚶(Scapharca broughtonii) Ets家族9条基因(分别命名为ETS-1~ ETS-9)，开放阅读框(ORF)大小分别为1065、1290、1569、912、1344、1404、1521、1968和1191 bp，并分别编码354、429、522、303、447、468、506、655和396个氨基酸。系统进化树分类表明，本研究所获得的基因均属Ets家族。对ETS-1和ETS-3的氨基酸序列和三维结构分析表明，其均含有高度保守的ETS结构域。在不同水温条件下，通过人工注射牡蛎疱疹病毒(OsHV-1)对魁蚶进行感染，并对病毒拷贝数和Ets的相对表达量进行定量分析。结果显示，ETS-1和ETS-3只在高温阳性组中相对表达量显著上调，与病毒拷贝数在高温条件下增长趋势呈正相关；ETS-4和ETS-8只在低温阳性组中相对表达量显著上调，但在高温阳性组中ETS-4和ETS-8的相对表达量与病毒拷贝数呈负相关；初步研究结果显示，从魁蚶Ets基因中筛选出2条Ets基因(ETS-1和ETS-3)，在高温条件下(16±2)℃，其可能参与正向调控病毒OsHV-1的复制过程。本研究为进一步探索魁蚶在夏季因感染牡蛎疱疹病毒(OsHV-1)而导致的大量死亡提供了科学数据。 |
| 关键词: 魁蚶 Ets家族 OsHV-1 克隆 温度 |
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| Cloning of Nine Genes in the Ets Family of Ark Clam (Scapharca broughtonii) and Their Expression in Response to Oyster Herpes Virus (OsHV-1) Infection |
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WEI Zhixin1,2, XIN Lusheng3,2, BAI Changming3,2, LI Yanan2, ZHANG Shumin2, LI Chenghua1, WANG Chongming3,2
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1.School of Marine Sciences, Ningbo University, Ningbo 315211;2.Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;3.Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071
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| Abstract: |
| Ets transcription-factor networks represent a model for how combinatorial gene expression is achieved. The characteristic feature of Ets factors is the conserved ETS domain (Helix-Turn-Helix). Thus, the Ets proteins bind to a core GGAA/T consensus sequence and regulate expression of several genes and play an important role in various cellular functions (mitosis, growth, development, differentiation, and apoptosis) and the regulation of immunity. In this experiment, 9 Ets genes (named ETS 1~ETS9, respectively) of the Ark clam (Scapharca broughtonii) were successfully obtained by gene cloning technology, and the open reading frames (ORFs) were 1065 bp, 1290 bp, 1569 bp, 912 bp, 1344 bp, 1404 bp, 1521 bp, 1968 bp, and 1191 bp, respectively. Moreover, they encoded respectively 354, 429, 522, 303, 447, 468, 506, 655, and 396 amino acids. Evolutionary relationships of taxa showed that all genes in this chapter belonged to the Ets family genes. The qPCR detection showed that the expression of two Ets genes (ETS-1, ETS-3) was significantly increased. Thus, the present study showed that the Ark clam ETS-1 and ETS-3 are involved in the replication process of the OsHV-1 under high temperature conditions (16±2)℃. In conclusion, the results of this study provide a scientific basis for further study concerning the large number of deaths of Ark clams due to infection with OsHV-1 in summer. |
| Key words: Scapharca broughtonii Ets family OsHV-1 Gene cloning Temperature |