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2019, 40(6): 121-130. doi: 10.19663/j.issn2095-9869.20190304001

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

1. 宁波大学海洋学院 宁波 315211;

2. 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;

3. 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

收稿日期:2019-03-04
修回日期:2019-03-11

基金项目:   现代农业产业体系专项资金(CARS-49)、中央级公益性科研院所基本科研业务费(20603022017007;20603022018014)共同资助 

关键词: 魁蚶 , Ets家族 , OsHV-1 , 克隆 , 温度

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

1. School of Marine Sciences, Ningbo University, Ningbo 315211;

2. Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266071;

3. 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

Received Date:2019-03-04
Accepted Date:2019-03-11

Keywords: Scapharca broughtonii , Ets family , OsHV-1 , Gene cloning , Temperature

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(Scapharcabroughtonii)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-1ETS-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-4ETS-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-4ETS-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-1ETS-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。

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.

参考文献

[1] Barbosa-Solomieu V, Dégremont L, Vázquez-Juárez R, et al. Ostreid herpesvirus 1 (OsHV-1) detection among three successive generations of Pacific oysters (Crassostrea gigas). Virus Research, 2005, 107(1):47-56
[2] Bosselut R, Duvall JF, Gégonne A, et al. The product of the c-ets-1 proto-oncogene and the related Ets2 protein act as transcriptional activators of the long terminal repeat of human T cell leukemia virus HTLV-1. Embo Journal, 1990, 9(10):3137-3144
[3] Elston R. Bivalve mollusc viruses. World Journal of Microbiology and Biotechnology, 1997, 13(4):393-403
[4] Graves BJ, Petersen JM. Specificity within the ets family of transcription factors. Advances in Cancer Research, 1998, 75:1-57
[5] Hine PM, Thorne T. A survey of some parasites and diseases of several species of bivalve mollusc in northern Western Australia. Diseases of Aquatic Organisms, 2000, 40(1):67-78
[6] 胡宗福, 任绍杰, 李树国. 牡蛎疱疹病毒(OsHV-1)研究进展. 水产科技情报, 2017, 44(5):259-264, 267Hu ZF, Ren SJ, Li SG. Research progress of oyster herpes virus (OsHV-1). Fisheries Science and Technology Information, 2017, 44(5):259-264, 267
[7] Jouaux A, Lafont M, Blin JL, et al. Physiological change under OsHV-1 contamination in Pacific oyster Crassostrea gigas through massive mortality events on fields. BMC Genomics, 2013, 14(1):590
[8] 李瑶瑶, 刘云国, 刘晓玲, 等. 基于线粒体基因组COI12S rRNA序列的魁蚶中国群体分类学地位研究. 渔业科学进展, 2018, 39(5):73-80Li YY, Liu YG, Liu XL, et al. The taxonomic status of Chinese population of Scapharca broughtonii based on sequences of COI and 12S rRNA genes. Progress in Fishery Sciences, 2018, 39(5):73-80
[9] 梁超, 杨爱国, 刘志鸿, 等. 魁蚶4个地理群体遗传结构的RAPD分析. 渔业科学进展, 2010, 31(1):59-64Liang C, Yang AG, Liu ZH, et al. RAPD analysis of genetic variations in four geographical populations of Scapharea broughtonii. Progress in Fishery Sciences, 2010, 31(1):59-64
[10] 刘寒苗, 吴彪, 刘志鸿, 等. 魁蚶(Scapharca broughtonii)不同地理群体的遗传多样性及种群结构. 渔业科学进展, 2017, 38(6):92-99Liu HM, Wu B, Liu ZL, et al. Genetic diversity and geographic population structures of Scapharca broughtonii. Progress in Fishery Sciences, 2017, 38(6):92-99
[11] 刘锡胤, 徐惠章. 魁蚶常温育苗技术. 科学养鱼, 2002(10):14-15Liu XY, Xu HZ. Seedling technique of Ark clam (Scapharca broughtonii) at room temperature. Sicentific Fish Faming, 2002(10):14-15
[12] Ma H, Wang J, Wang B, et al. Characterization of an ETS transcription factor in the sea scallop Chlamys farreri. Developmental and Comparative Immunology, 2009, 33(9):953-958
[13] 毛雪英, 阎子娟, 邵雁群. 魁蚶人工育苗及保苗的新探索. 河北渔业, 2007(4):38-39Mao XY, Yan ZJ, Shao YQ. A new exploration of artificial seedling and seedling preservation in Ark clams (Scapharca broughtonii). Hebei Fisheries, 2007(4):38-39
[14] Oikawa T, Yamada T. Molecular biology of the Ets family of transcription factors. Gene, 2003, 303(1-2):11-34
[15] Panagoulias I, Karagiannis F, Aggeletopoulou I, et al. Ets-2 acts as a transcriptional repressor of the human immunodeficiency virus type 1 through binding to a repressor-Activator target sequence of 5'-LTR. Frontiers Immunology, 2018, 8:1924
[16] Pleschka S, Wolff T, Ehrhardt C, et al. Influenza virus propagation is impaired by inhibition of the RAF/MEK/ERK signalling cascade. Nature Cell Biology, 2001, 3(3):301-305
[17] Ren WC, Chen HX, Renault T, et al. Complete genome sequence of acute viral necrosis virus associated with massive mortality outbreaks in the Chinese scallop, Chlamys farreri. Virology Journal, 2013, 10:110
[18] Renault T, Lipart C, Arzul I. A herpes-like virus infecting Crassostrea gigas and Ruditapes philippinarum larvae in France. Journal of Fish Diseases, 2001, 24(6):369-376
[19] Roque A, Carrasco N, Andree KB, et al. First report of OsHV-1 microvar in Pacific oyster (Crassostrea gigas) cultured in Spain. Aquaculture, 2012, 324(3):303-306
[20] Ruelas D, Greene W. An integrated overview of HIV-1 latency. Cell, 2013, 155(3):519-529
[21] Segarra A, Pépin JF, Arzul I, et al. Detection and description of a particular Ostreid herpesvirus 1 genotype associated with massive mortality outbreaks of Pacific oysters, Crassostrea gigas, in France in 2008. Virus Research, 2010, 153(1):92-99
[22] Seth A, Hodge DR, Thompson DM, et al. ETS family proteins activate transcription from HIV-1 long terminal repeat. Aids Research and Human Retroviruses, 1993, 9(19):1017-1023
[23] Sharrocks AD, Brown AL, Ling Y, et al. The ETS-domain transcription factor family. International Journal of Biochemistry and Cell Biology, 1997, 29(12):1371-1387
[24] Sieweke MH, Tekotte H, Jarosch U, et al. Cooperative interaction of Ets-1 with USF-1 required for HIV-1 enhancer activity in T cells. Embo Journal, 1998, 17(6):1728-1739
[25] Suico MA, Shuto T, Kai H. Roles and regulations of the ETS transcription factor ELF4/MEF. Journal of Molecular Cell Biology, 2017, 9(3):168-177
[26] Wang B, Krall EB, Aguirre AJ, et al. ATXN1L, CIC, and ETS transcription factors modulate sensitivity to MAPK pathway inhibition. Cell Reports, 2017, 18(6):1543-1557
[27] Wasylyk B, Hagman J, Gutierrez-Hartmann A. Ets transcription
[28] factors:Nuclear effectors of the Ras-MAP-kinase signaling pathway. Trends in Biochemical Science, 1998, 23(6):213-216
[29] Webb SC, Fidler A, Renault T, et al. Primers for PCR-based detection of ostreid herpes virus-1 (OsHV-1):Application in a survey of New Zealand molluscs. Aquaculture, 2007, 272(1-4):126-139
[30] Xia JY, Bai CM, Wang CM, et al. Complete genome sequence of Ostreid herpesvirus-1 associated with mortalities of Scapharca broughtonii broodstocks. Virology Journal, 2015, 12:110
[31] Xin LS, Huang BW, Bai CM, et al. Validation of housekeeping genes for quantitative mRNA expression analysis in OsHV-1 infected Ark clam, Scapharca broughtonii. Journal of Invertebrate Pathology, 2018, 155:44-51
[32] 曾红, 杨洋, 安输, 等. Ets转录因子的生理作用研究进展. 中国药理学通报, 2017, 33(12):1645-1650Zeng H, Yang Y, An Y, et al. Physiogical functions of Ets family of transcription factors. China Pharmacological Bulletin, 2017, 33(12):1645-1650
[33] 张启刚, 王如才, 王兴章, 等. 魁蚶(Arca (Anadara) inflata Reeve)品系选育. 现代渔业信息, 2007, 22(7):25-26Zhang QG, Wang RC, Wang XZ, et al. Selective breeding of strain of Arca (Anadara) inflate reeve. Modern Fisheries Information, 2007, 22(7):25-26

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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