牙鲆efhd2和tbc1d25基因的克隆和表达分析
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摘要
为在分子水平解析牙鲆(Paralichthys olivaceus)抗淋巴囊肿病的机理,本研究克隆牙鲆淋巴囊肿抗病免疫相关基因efhd2和tbc1d25的c DNA全序列,运用生物信息学方法对基因序列进行了详细分析;同时,采用荧光定量PCR分析了efhd2和tbc1d25基因在牙鲆胚胎发育不同阶段、淋巴囊肿抗病和患病个体不同组织的表达特征。结果显示,efhd2基因c DNA全长为5231 bp,开放阅读框(ORF)长为699 bp,编码232个氨基酸。tbc1d25基因c DNA全长为3173 bp,ORF长为2601 bp,编码866个氨基酸。定量结果显示,efhd2和tbc1d25基因在胚胎发育的各个时期均有不同程度的表达,其中,efhd2在出膜仔鱼期表达量最高,而tbc1d25在受精卵中的表达量显著高于其他时期(P<0.05)。在所研究的淋巴囊肿抗病和患病个体不同组织中,efhd2和tbc1d25基因均有不同程度的表达,抗病个体的血液中,这2个基因的表达量均显著高于患病个体(P<0.05)。本研究结果为深入探讨efhd2和tbc1d25基因功能和解析牙鲆淋巴囊肿抗病机理提供了基础资料。
The Japanese flounder(Paralichthys olivaceus) is one of the most important marine culture species in China. However, the outbreak of diseases has seriously affected the industrial culture of this particular fish. Among these diseases, the one caused by the lymphocystis disease virus has been spreading widely and has resulted in severe economic losses every year. In order to select new varieties of Japanese flounder that are resistant to lymphocystis disease in China, and to elucidate the mechanism of disease resistance at the molecular level, we used a high-throughput sequencing technique to analyze the transcriptome of kidney tissues of the Japanese flounder, and screened out a number of functional genes closely related to resistance, including efhd2 and tbc1 d25. In this study, we cloned the full-length cDNA sequences of efhd2 and tbc1 d25 by using RACE(rapid-amplification of cDNA ends). The efhd2 gene was5231 bp in length, of which the length of the 5' untranslated region(5' UTR) was 142 bp and the length of the 3' UTR was 4390 bp. The open reading frame(ORF) was 699 bp in length, and encoded 232 amino acids with a molecular weight of 26.4 kDa and an isoelectric point of about 5.08. The full length of the tbc1 d25 gene was 3173 bp, of which the 5' UTR length was 108 bp, and the 3' UTR length was 464 bp.The ORF was 2601 bp in length and encoded 866 amino acids with a molecular weight of 96.4 kDa and an isoelectric point of about 5.47. Multiple sequence alignment revealed that the amino acid sequences of EFHD2 have 83%, 88%, 73%, and 72% homology with Danio rerio, Oryzias latipes, Mus musculus, and Homo sapiens respectively; and those of TBC1 D25 have 71%, 74%, 72% and 74% homology with these four species, respectively. The expression profiles of efhd2 and tbc1 d25 were analyzed by quantitative real-time PCR(qRT-PCR). Both efhd2 and tbc1 d25 were expressed at the fertilized egg, 4-cell, 32-cell,128-cell, high blastocyst, low blastocyst, early gastrula, late gastrula, sarcomere, heartbeat, and hatched larva stages. The expression level of efhd2 in the 4-cell, low blastocyst and early gastrula stage was lower than that in other stages(P<0.05); the expression level of efhd2 began to increase from the sarcomere stage and reached the highest level at the hatched larva stage, and was significantly higher than that in other groups(P<0.05). The expression level of the tbc1 d25 gene in the fertilized egg was significantly higher than that in other stages(P<0.05); during development, the expression level of tbc1 d25 decreased to the lowest level in late gastrula(P<0.05); there was no significant difference in the expression level of tbc1 d25 in late gastrula, sarcomere, and heartbeat stages(P<0.05). In the lymphocystis disease-resistant and-sensitive individuals, the expression of efhd2 and tbc1 d25 was detected in head, kidneys, liver, blood,gills, heart, gonads, muscle, intestine and spleen. The expression of efhd2 and tbc1 d25 was significantly higher in the blood of lymphocystis disease-resistant individuals than in the lymphocystis disease-sensitive individuals. This study provides a basis for further studying the gene function of efhd2 and tbc1 d25, as well as the mechanism of disease resistance of the Japanese flounder to lymphocystis disease.
The Japanese flounder(Paralichthys olivaceus) is one of the most important marine culture species in China. However, the outbreak of diseases has seriously affected the industrial culture of this particular fish. Among these diseases, the one caused by the lymphocystis disease virus has been spreading widely and has resulted in severe economic losses every year. In order to select new varieties of Japanese flounder that are resistant to lymphocystis disease in China, and to elucidate the mechanism of disease resistance at the molecular level, we used a high-throughput sequencing technique to analyze the transcriptome of kidney tissues of the Japanese flounder, and screened out a number of functional genes closely related to resistance, including efhd2 and tbc1 d25. In this study, we cloned the full-length cDNA sequences of efhd2 and tbc1 d25 by using RACE(rapid-amplification of cDNA ends). The efhd2 gene was5231 bp in length, of which the length of the 5' untranslated region(5' UTR) was 142 bp and the length of the 3' UTR was 4390 bp. The open reading frame(ORF) was 699 bp in length, and encoded 232 amino acids with a molecular weight of 26.4 kDa and an isoelectric point of about 5.08. The full length of the tbc1 d25 gene was 3173 bp, of which the 5' UTR length was 108 bp, and the 3' UTR length was 464 bp.The ORF was 2601 bp in length and encoded 866 amino acids with a molecular weight of 96.4 kDa and an isoelectric point of about 5.47. Multiple sequence alignment revealed that the amino acid sequences of EFHD2 have 83%, 88%, 73%, and 72% homology with Danio rerio, Oryzias latipes, Mus musculus, and Homo sapiens respectively; and those of TBC1 D25 have 71%, 74%, 72% and 74% homology with these four species, respectively. The expression profiles of efhd2 and tbc1 d25 were analyzed by quantitative real-time PCR(qRT-PCR). Both efhd2 and tbc1 d25 were expressed at the fertilized egg, 4-cell, 32-cell,128-cell, high blastocyst, low blastocyst, early gastrula, late gastrula, sarcomere, heartbeat, and hatched larva stages. The expression level of efhd2 in the 4-cell, low blastocyst and early gastrula stage was lower than that in other stages(P<0.05); the expression level of efhd2 began to increase from the sarcomere stage and reached the highest level at the hatched larva stage, and was significantly higher than that in other groups(P<0.05). The expression level of the tbc1 d25 gene in the fertilized egg was significantly higher than that in other stages(P<0.05); during development, the expression level of tbc1 d25 decreased to the lowest level in late gastrula(P<0.05); there was no significant difference in the expression level of tbc1 d25 in late gastrula, sarcomere, and heartbeat stages(P<0.05). In the lymphocystis disease-resistant and-sensitive individuals, the expression of efhd2 and tbc1 d25 was detected in head, kidneys, liver, blood,gills, heart, gonads, muscle, intestine and spleen. The expression of efhd2 and tbc1 d25 was significantly higher in the blood of lymphocystis disease-resistant individuals than in the lymphocystis disease-sensitive individuals. This study provides a basis for further studying the gene function of efhd2 and tbc1 d25, as well as the mechanism of disease resistance of the Japanese flounder to lymphocystis disease.
引文
Anders K.Lymphocystis disease of fishes,in viruses of lower vertebrates.Heidelberg:Springer,1989,141-160
Avramidou A,Kroczek C,Lang C,et al.The novel adaptor protein Swiprosin-1 enhances BCR signals and contributes to BCR-induced apoptosis.Cell Death and Differentiation,2007,14(11):1936-1947
Brachs S,Lang C,Buslei R,et al.Monoclonal antibodies to discriminate the ef hand containing calcium binding adaptor proteins EFhd1 and EFhd2.Monoclonal Antibodies in Immunodiagnosis and Immunotherapy,2013,32(4):237-245
Castillo A,Razquin B,Villena AJ,et al.Thymic barriers to antigen entry during the post-hatching development of the thymus of rainbow trout,Oncorhynchus mykiss.Fish&Shellfish Immunology,1998,8(3):157-170
Chen Z,Jin T,Lu Y.Antimi R-30b inhibits TNF-αmediated apoptosis and attenuated cartilage degradation through enhancing autophagy.Cellular Physiology and Biochemistry2016,40(5):883-894
Corona AK,Jackson WT.Finding the middle ground for autophagic fusion requirements.Trends in Cell Biology,2018,28(11):869-881
Dalmo RA,Ingebrigtsen K,B?gwald J.Non-specific defense mechanisms in fish,with particular reference to the reticuloendothelial system(RES).Journal of Fish Diseases,1997,20(4):241-273
Fan CX,Chen SL,Wang L,et al.Screening and identification of SSR markers associated with lymphocystis disease resistance in Japanese flounder(Paralichthys olivaceus).Journal of Fisheries of China,2014,38(4):577-583[范彩霞,陈松林,王磊,等.牙鲆抗淋巴囊肿病相关SSR标记的筛选与鉴定.水产学报,2014,38(4):577-583]
Frasa MAM,Koessmeier KT,Ahmadian MR,et al.Illuminating the functional and structural repertoire of human TBC/RABGAPs.Nature Reviews Molecular Cell Biology,2012,13(2):67-73
Fuji K,Kobayashi K,Hasegawa O,et al.Identification of a single major genetic locus controlling the resistance to lymphocystis disease in Japanese flounder(Paralichthys olivaceus).Aquaculture,2006,254(1):203-210
Fukuda M.Membrane traffic in the secretory pathway.Cellular and Molecular Life Sciences,2008,65(18):2801-2813
Fukuda M.TBC proteins:GAPs for mammalian small GTPase Rab?Bioscience Reports,2011,31(3):159-168
Gorski SM,Chittaranjan S,Pleasance ED,et al.A SAGEapproach to discovery of genes involved in autophagic cell death.Current Biology,2003,13(4):358-363
Grosshans BL,Ortiz D,Novick P.Rabs and their effectors:Achieving specificity in membrane traffic.Proceedings of the National Academy of Sciences,2006,103(32):11821-11827
Hassan M,Selimovic D,Hannig M,et al.Endoplasmic reticulum stress-mediated pathways to both apoptosis and autophagy:Significance for melanoma treatment.World Journal of Experimental Medicine,2015,5(4):206-217
Hou JL,Wang GX,Zhang XY,et al.Analysis of growth and lymphocystis disease resistance of selected breeding families of Japanese flounder(Paralichthys olivaceus).Journal of Fishery Sciences of China,2017,24(4):727-737[侯吉伦,王桂兴,张晓彦,等.牙鲆抗淋巴囊肿病家系选育及生长和抗病性能分析.中国水产科学,2017,24(4):727-737]
Hwang SD,Fuji K,Takano T,et al.Linkage mapping of Toll-like receptors(TLRs)in Japanese flounder,Paralichthys olivaceus.Marine Biotechnology,2011,13(6):1086-1091
Itoh T,Fukuda M.Roles of Rab-GAPs in regulating autophagy,in autophagy:Cancer,other pathologies,inflammation,immunity,infection,and aging.Cambridge:Academy Press,2017,143-157
Itoh T,Kanno E,Uemura T,et al.OATL1,a novel autophagosomeresident Rab33B-GAP,regulates autophagosomal maturation.Journal of Cell Biology,2011,192(5):839-853
Kim YD,Kwon MS,Na BR,et al.Swiprosin-1 expression is up-regulated through protein kinase C-θand NF-κBpathway in T cells.Immune Network,2013,13(2):55-62
Kinne O.Diseases of marine animals.Volume IV,Part 1,Introduction,Pisces.Hamburg:Biologische Anstalt Helgoland1984,1-541
Kroczek C,Lang C,Brachs S,et al.Swiprosin-1/EFhd2 controls B cell receptor signaling through the assembly of the B cell receptor,Syk,and phospholipase C2 in membrane rafts.Journal of Immunology,2010,184(7):3665-3676
Liu HJ,Wang CA,Zhu XC,et al.Embryonic development of gynogenetic diploid and triploid Japanese flounder Paralichthys olivaceus.Journal of Dalian Fisheries University,2008,23(3):161-167[刘海金,王常安,朱晓琛,等.牙鲆单倍体,三倍体,雌核发育二倍体和普通二倍体胚胎发育的比较.大连水产学院学报,2008,23(3):161-167]
Ma HX,Wang L,Wang LQ,et al.The different expression patterns of the gene Arginase II in Paralichthys olivaceus and the correlation between its expression and Edwardsiella tarda infection.Progress in Fishery Sciences,2018,39(3):44-52[马慧鑫,王磊,汪林庆,等.牙鲆精氨酸酶Ⅱ基因的克隆以及免疫应答表达分析.渔业科学进展,2018,39(3):44-52]
Paperna I,Vilenkin M,Alves de Matos AP.Iridovirus infections in farm-reared tropical ornamental fish.Diseases of Aquatic Organisms,2001,48(1):17-25
Pereira-Leal JB,Seabra MC.Evolution of the rab family of small GTP-binding proteins.Journal of Molecular Biology,2001,313(4):889-901
Qu LY,Sun XQ,Zhang JX.Electron ultramicroscopic observation on lymphocystis disease of the left-eyed flounder Paralichthys olivaceus.Journal of Qingdao Ocean University,2000,30(2):105-110[曲凌云,孙修勤,张进兴.养殖牙鲆淋巴囊肿病的电镜观察.青岛海洋大学学报,2000,30(2):105-110]
Schwartz SL,Cao C,Pylypenko O,et al.Rab GTPases at a glance.Journal of Cell Science,2007,120(22):3905-3910
Shao CW,Bao BL,Xie ZY,et al.The genome and transcriptome of Japanese flounder provide insights into flatfish asymmetry.Nature Genetics,2017,49(1):119-124
Si F,Sun ZH,Yu SS,et al.Analysis of the genetic variability of the captured population of Japanese flounder Paralichthys olivaceus in the coastal releasing area of Qinhuangdao.Progress in Fishery Sciences,2017,38(6):18-24[司飞,孙朝徽,于姗姗,等.秦皇岛近海增殖放流区牙鲆(Paralichthys olivaceus)回捕群体的遗传多样性分析.渔业科学进展,2017,38(6):18-24]
Stenmark H.Rab GTPases as coordinators of vesicle traffic.Nature Reviews Molecular Cell Biology,2009,10(8):513-525
Vega IE.EFhd2,a protein linked to alzheimer’s disease and other neurological disorders.Frontiers in Neuroscience,2016,10:150
Vuadens F,Rufer N,Kress A,et al.Identification of swiprosin 1in human lymphocytes.Proteomics,2004,4(8):2216-2220
Wei XL,Xu ZC,Wang GX,et al.pBACode:A random-barcodebased high-throughput approach for BAC paired-end sequencing and physical clone mapping.Nucleic Acids Research,2017,45(7):e52
Wolf K.Fish viruses and fish viral diseases.New York:Cornell University Press,1988,476
Wu R,Sheng X,Tang X,et al.Transcriptome analysis of flounder(Paralichthys olivaceus)gill in response to lymphocystis disease virus(LCDV)infection:Novel insights into fish defense mechanisms.International Journal of Molecular Sciences,2018,19(1):160
Wu R,Tang X,Sheng X,et al.Relationship between expression of cellular receptor-27.8kDa and lymphocystis disease virus(LCDV)infection.PLoS One,2015,10(5):e0127940
Yang Y,Huang YH,Yu YP,et al.RING domain is essential for the antiviral activity of TRIM25 from orange spotted grouper.Fish&Shellfish Immunology,2016,55:304-314
Young ARJ,Chan EYW,Hu XW,et al.Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes.Journal of Cell Science,2006,119(18):3888-3900
Zeng R,Chen Y,Zhao S,et al.Autophagy counteracts apoptosis in human multiple myeloma cells exposed to oridonin in vitro via regulating intracellular ROS and SIRT1.Acta Pharmacologica Sinica,2012,33(1):91-100
Zhang WT,Xiang JS,Li HL,et al.Molecular cloning,characterization,and expression of three TLR5S splicing variants in half-smooth tongue sole.Journal of Fishery Sciences of China,2016,23(1):10-20[张文婷,向晋松,李海龙,等.半滑舌鳎TLR5S三种剪切型基因的克隆与表达分析.中国水产科学,2016,23(1):10-20]
Zheng FR,Sun XQ,Xing MQ,et al.Immune response of DNAvaccine against lymphocystis disease virus and expression analysis of immune-related genes after vaccination.Aquaculture Research,2010,41(10):1444-1451
Zheng JM,Gao FY,Lu MX,et al.Cloning and expression analysis of TRIM16 and TRIM25 genes from Nile Tilapia(Oreochromis niloticus).Journal of Agricultural Biotechnology,2017,25(6):861-873[郑建美,高凤英,卢迈新,等.尼罗罗非鱼TRIM16和TRIM25基因的克隆及表达分析.农业生物技术学报,2017,25(6):861-873]
Avramidou A,Kroczek C,Lang C,et al.The novel adaptor protein Swiprosin-1 enhances BCR signals and contributes to BCR-induced apoptosis.Cell Death and Differentiation,2007,14(11):1936-1947
Brachs S,Lang C,Buslei R,et al.Monoclonal antibodies to discriminate the ef hand containing calcium binding adaptor proteins EFhd1 and EFhd2.Monoclonal Antibodies in Immunodiagnosis and Immunotherapy,2013,32(4):237-245
Castillo A,Razquin B,Villena AJ,et al.Thymic barriers to antigen entry during the post-hatching development of the thymus of rainbow trout,Oncorhynchus mykiss.Fish&Shellfish Immunology,1998,8(3):157-170
Chen Z,Jin T,Lu Y.Antimi R-30b inhibits TNF-αmediated apoptosis and attenuated cartilage degradation through enhancing autophagy.Cellular Physiology and Biochemistry2016,40(5):883-894
Corona AK,Jackson WT.Finding the middle ground for autophagic fusion requirements.Trends in Cell Biology,2018,28(11):869-881
Dalmo RA,Ingebrigtsen K,B?gwald J.Non-specific defense mechanisms in fish,with particular reference to the reticuloendothelial system(RES).Journal of Fish Diseases,1997,20(4):241-273
Fan CX,Chen SL,Wang L,et al.Screening and identification of SSR markers associated with lymphocystis disease resistance in Japanese flounder(Paralichthys olivaceus).Journal of Fisheries of China,2014,38(4):577-583[范彩霞,陈松林,王磊,等.牙鲆抗淋巴囊肿病相关SSR标记的筛选与鉴定.水产学报,2014,38(4):577-583]
Frasa MAM,Koessmeier KT,Ahmadian MR,et al.Illuminating the functional and structural repertoire of human TBC/RABGAPs.Nature Reviews Molecular Cell Biology,2012,13(2):67-73
Fuji K,Kobayashi K,Hasegawa O,et al.Identification of a single major genetic locus controlling the resistance to lymphocystis disease in Japanese flounder(Paralichthys olivaceus).Aquaculture,2006,254(1):203-210
Fukuda M.Membrane traffic in the secretory pathway.Cellular and Molecular Life Sciences,2008,65(18):2801-2813
Fukuda M.TBC proteins:GAPs for mammalian small GTPase Rab?Bioscience Reports,2011,31(3):159-168
Gorski SM,Chittaranjan S,Pleasance ED,et al.A SAGEapproach to discovery of genes involved in autophagic cell death.Current Biology,2003,13(4):358-363
Grosshans BL,Ortiz D,Novick P.Rabs and their effectors:Achieving specificity in membrane traffic.Proceedings of the National Academy of Sciences,2006,103(32):11821-11827
Hassan M,Selimovic D,Hannig M,et al.Endoplasmic reticulum stress-mediated pathways to both apoptosis and autophagy:Significance for melanoma treatment.World Journal of Experimental Medicine,2015,5(4):206-217
Hou JL,Wang GX,Zhang XY,et al.Analysis of growth and lymphocystis disease resistance of selected breeding families of Japanese flounder(Paralichthys olivaceus).Journal of Fishery Sciences of China,2017,24(4):727-737[侯吉伦,王桂兴,张晓彦,等.牙鲆抗淋巴囊肿病家系选育及生长和抗病性能分析.中国水产科学,2017,24(4):727-737]
Hwang SD,Fuji K,Takano T,et al.Linkage mapping of Toll-like receptors(TLRs)in Japanese flounder,Paralichthys olivaceus.Marine Biotechnology,2011,13(6):1086-1091
Itoh T,Fukuda M.Roles of Rab-GAPs in regulating autophagy,in autophagy:Cancer,other pathologies,inflammation,immunity,infection,and aging.Cambridge:Academy Press,2017,143-157
Itoh T,Kanno E,Uemura T,et al.OATL1,a novel autophagosomeresident Rab33B-GAP,regulates autophagosomal maturation.Journal of Cell Biology,2011,192(5):839-853
Kim YD,Kwon MS,Na BR,et al.Swiprosin-1 expression is up-regulated through protein kinase C-θand NF-κBpathway in T cells.Immune Network,2013,13(2):55-62
Kinne O.Diseases of marine animals.Volume IV,Part 1,Introduction,Pisces.Hamburg:Biologische Anstalt Helgoland1984,1-541
Kroczek C,Lang C,Brachs S,et al.Swiprosin-1/EFhd2 controls B cell receptor signaling through the assembly of the B cell receptor,Syk,and phospholipase C2 in membrane rafts.Journal of Immunology,2010,184(7):3665-3676
Liu HJ,Wang CA,Zhu XC,et al.Embryonic development of gynogenetic diploid and triploid Japanese flounder Paralichthys olivaceus.Journal of Dalian Fisheries University,2008,23(3):161-167[刘海金,王常安,朱晓琛,等.牙鲆单倍体,三倍体,雌核发育二倍体和普通二倍体胚胎发育的比较.大连水产学院学报,2008,23(3):161-167]
Ma HX,Wang L,Wang LQ,et al.The different expression patterns of the gene Arginase II in Paralichthys olivaceus and the correlation between its expression and Edwardsiella tarda infection.Progress in Fishery Sciences,2018,39(3):44-52[马慧鑫,王磊,汪林庆,等.牙鲆精氨酸酶Ⅱ基因的克隆以及免疫应答表达分析.渔业科学进展,2018,39(3):44-52]
Paperna I,Vilenkin M,Alves de Matos AP.Iridovirus infections in farm-reared tropical ornamental fish.Diseases of Aquatic Organisms,2001,48(1):17-25
Pereira-Leal JB,Seabra MC.Evolution of the rab family of small GTP-binding proteins.Journal of Molecular Biology,2001,313(4):889-901
Qu LY,Sun XQ,Zhang JX.Electron ultramicroscopic observation on lymphocystis disease of the left-eyed flounder Paralichthys olivaceus.Journal of Qingdao Ocean University,2000,30(2):105-110[曲凌云,孙修勤,张进兴.养殖牙鲆淋巴囊肿病的电镜观察.青岛海洋大学学报,2000,30(2):105-110]
Schwartz SL,Cao C,Pylypenko O,et al.Rab GTPases at a glance.Journal of Cell Science,2007,120(22):3905-3910
Shao CW,Bao BL,Xie ZY,et al.The genome and transcriptome of Japanese flounder provide insights into flatfish asymmetry.Nature Genetics,2017,49(1):119-124
Si F,Sun ZH,Yu SS,et al.Analysis of the genetic variability of the captured population of Japanese flounder Paralichthys olivaceus in the coastal releasing area of Qinhuangdao.Progress in Fishery Sciences,2017,38(6):18-24[司飞,孙朝徽,于姗姗,等.秦皇岛近海增殖放流区牙鲆(Paralichthys olivaceus)回捕群体的遗传多样性分析.渔业科学进展,2017,38(6):18-24]
Stenmark H.Rab GTPases as coordinators of vesicle traffic.Nature Reviews Molecular Cell Biology,2009,10(8):513-525
Vega IE.EFhd2,a protein linked to alzheimer’s disease and other neurological disorders.Frontiers in Neuroscience,2016,10:150
Vuadens F,Rufer N,Kress A,et al.Identification of swiprosin 1in human lymphocytes.Proteomics,2004,4(8):2216-2220
Wei XL,Xu ZC,Wang GX,et al.pBACode:A random-barcodebased high-throughput approach for BAC paired-end sequencing and physical clone mapping.Nucleic Acids Research,2017,45(7):e52
Wolf K.Fish viruses and fish viral diseases.New York:Cornell University Press,1988,476
Wu R,Sheng X,Tang X,et al.Transcriptome analysis of flounder(Paralichthys olivaceus)gill in response to lymphocystis disease virus(LCDV)infection:Novel insights into fish defense mechanisms.International Journal of Molecular Sciences,2018,19(1):160
Wu R,Tang X,Sheng X,et al.Relationship between expression of cellular receptor-27.8kDa and lymphocystis disease virus(LCDV)infection.PLoS One,2015,10(5):e0127940
Yang Y,Huang YH,Yu YP,et al.RING domain is essential for the antiviral activity of TRIM25 from orange spotted grouper.Fish&Shellfish Immunology,2016,55:304-314
Young ARJ,Chan EYW,Hu XW,et al.Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes.Journal of Cell Science,2006,119(18):3888-3900
Zeng R,Chen Y,Zhao S,et al.Autophagy counteracts apoptosis in human multiple myeloma cells exposed to oridonin in vitro via regulating intracellular ROS and SIRT1.Acta Pharmacologica Sinica,2012,33(1):91-100
Zhang WT,Xiang JS,Li HL,et al.Molecular cloning,characterization,and expression of three TLR5S splicing variants in half-smooth tongue sole.Journal of Fishery Sciences of China,2016,23(1):10-20[张文婷,向晋松,李海龙,等.半滑舌鳎TLR5S三种剪切型基因的克隆与表达分析.中国水产科学,2016,23(1):10-20]
Zheng FR,Sun XQ,Xing MQ,et al.Immune response of DNAvaccine against lymphocystis disease virus and expression analysis of immune-related genes after vaccination.Aquaculture Research,2010,41(10):1444-1451
Zheng JM,Gao FY,Lu MX,et al.Cloning and expression analysis of TRIM16 and TRIM25 genes from Nile Tilapia(Oreochromis niloticus).Journal of Agricultural Biotechnology,2017,25(6):861-873[郑建美,高凤英,卢迈新,等.尼罗罗非鱼TRIM16和TRIM25基因的克隆及表达分析.农业生物技术学报,2017,25(6):861-873]