| 221 | 5 | 49 |
| 下载次数 | 被引频次 | 阅读次数 |
超氧化物歧化酶(SOD)是通过歧化反应催化超氧化物转化为O2和H2O2的酶,铜锌超氧化物歧化酶(CuZnSOD)和锰超氧化物歧化酶(MnSOD)是两种主要存在形式。本研究通过RACE法和RT-qPCR技术对布氏鲳鲹(Trachinotus blochii)的SOD-1、SOD-2进行基因克隆、序列分析和组织分布的研究。结果显示,布氏鲳鲹SOD-1 (icCuZnSOD)基因的ORF长486 bp,编码161个氨基酸,分子量为16.676 kD,蛋白结构预测无信号肽,含有1个SOD-CU结构域。布氏鲳鲹SOD-2 (mMnSOD)基因的ORF长657 bp,编码218个氨基酸,分子量为24.261 k D,蛋白结构预测无信号肽,含有1个SOD-FE蛋白结构域。氨基酸序列同源比对结果表明,布氏鲳鲹SOD-1有1段与其他鱼类显著差异的精氨酸-天冬酰胺-丝氨酸-丙氨酸-半胱氨酸-谷氨酰胺-异亮氨酸序列,且与鲈形目点带石斑鱼(Epinephelus coioides)、斜带石斑鱼(Epinephelus malabaricus)CuZnSOD同源性较高;SOD-2与鲈形目的红甘鲹(Seriola dumerili) MnSOD同源性较高且聚为一支。组织分布结果表明,SOD-1和SOD-2在布氏鲳鲹的心脏和肝脏中表达量最高,其次在脑和鳃表达量相对较高,表明SOD-1和SOD-2可能在免疫反应和维持新陈代谢方面有重要作用。本研究为今后布氏鲳鲹的抗逆、免疫的研究提供了研究依据。
Abstract：Superoxide dismutases(SOD) are some enzymes that catalyze superoxide to oxygen(O2) and hydrogen peroxide(H_2O2) by disproportionation. There are two types of CuZnSOD and MnSOD in cells. In this study, the cloning and tissue distribution of icCuZnSOD(SOD-1) and(SOD-2) in Trachinotus blochii were studied by RACE and RT-qPCR. The studies showed that the ORF of SOD-1(icCuZnSOD) was 486 bp and encoded 161 amino acids with a molecular weight of 16.676 kD, and SOD-1 had no signal peptide but had a SOD-CU protein domain.The ORF of SOD-2(mMnSOD) was 657 bp and encoded 218 amino acids with a molecular weight of 24.261 kD,and SOD-2 also had no signal peptide with a SOD-FE protein domain. By homologous alignment of amino acid sequences, a different peptide fragment was found in SOD-1 that contains Arg-Asn-Ser-Ala-Cys-Gln-Ile. It was found that SOD-1 was closely related to Epinephelus coioides and Epinephelus malabaricus, and SOD-2 was closely related to Seriola dumerili and forms its own branch. Real-time quantitative PCR showed that the expression of SOD-1 and SOD-2 was the highest in the heart and liver, followed by relatively high expression in the brain and gill. It indicated that SOD-1 and SOD-2 may be involved in immune epidemic prevention and maintaining metabolism. This study lays a theoretical foundation for the study of the stress resistance and immune of Trachinostus blochii in the future
Anju A.,Jeswin J.,Thomas P.C.,Paulton M.P.,and Vijayan K.,2013,Molecular cloning,characterization and expression analysis of cytoplasmic Cu/Zn-superoxide dismutase(SOD)from pearl oyster Pinctada fucata,Fish Shellfish Immunol.,34(3):946-950.
Bao Y.,Li L.,Xu F.,and Zhang G.,2009,Intracellular copper/zinc superoxide dismutase from bay scallop Argopecten irradians:Its gene structure,m RNA expression and recombinant protein,Fish Shellfish Immunol.,27(2):210-220.
Beutler B.,2004,Innate immunity:an overview,Molecular Immunology,40(12):845-859.
Chakravarthy N.,Aravindan K.,Kalaimani N.,Alavandi S.V.,Poornima M.,and Santiago T.C.,2012,Intracellular copper zinc superoxide dismutase(icCuZnSOD) from Asian seabass(Lates calcarifer):molecular cloning,characterization and gene expression with reference to Vibrio anguillarum infection,Developmental&Comparative Immunology,36(4):751-755.
Chen Q.L.,Sun Y.L.,Liu Z.H.,and Li Y.W.,2017,Sex-dependent effects of subacute mercuric chloride exposure on histology,antioxidant status and immune-related gene expression in the liver of adult zebrafish(Danio rerio),Chemosphere,188:1-9.
Cheng W.,Tung Y.H.,Chiou T.T.,and Chen J.C.,2006,Cloning and characterisation of mitochondrial manganese superoxide dismutase(mtMnSOD) from the giant freshwater prawn Macrobrachium rosenbergii,Fish Shellfish Immunol.,21(4):453-466.
Cheng Z.Y.,Xu L.Q.,Hu X.P.,Hu B.Q.,Jian S.Q.,Yang G.,Zhang M.,Wen C.G.,Ning R.H.,and Wang H.,2015,Cloning and prokaryotic expression of copper/zinc superoxide dismutase gene from Siniperca chuatsi,Nanchang Daxue Xuebao(Journal of Nanchang University(Natural Science)),39(6):587-596.(程周玉,许亮清,胡向萍,胡宝庆,简少卿,阳刚,张明,文春根,宁瑞红,王鹤,2015,翘嘴鳜铜锌超氧化物歧化酶基因的分子克隆与原核表达,南昌大学学报(理科版),39(6):587-596.)
Gao X.G.,He C.B.,Liu H.,Li H.J.,Zhu D.,Cai S.L.,Xia Y.,Wang Y.,and Yu Z.,2012,Intracellular Cu/Zn superoxide dismutase(Cu/Zn-SOD) from hard clam Meretrix meretrix:its cDNA cloning,m RNA expression and enzyme activity,Molecular Biology Reports,39(12):10713-10722.
Jesu A.,Rajesh P.,Prasanth B.,Venkatesh K.,Gnanam A.J.,Mukesh P.,and Marimuthu K.,2014,Anovel murrel Channa striatus mitochondrial manganese superoxide dismutase:gene silencing,SOD activity,superoxide anion production and expression,Fish Physiology&Biochemistry,40(6):1937-1955.
Keilin D.,and Mann T.,1938,Polyphenol oxidase purification,nature and properties,Proceedings of the Royal Society of London,125(839):187-204.
Ken C.F.,Weng D.F.,Duan K.J.,and Lin C.T.,2002,Characterization of copper/zinc-superoxide dismutase from Pagrus major cDNA and enzyme stability,Journal of Agricultural&Food Chemistry,50(4):784.
Liu H.,He J.Y.,Chi C.F.,and Gu Y.F.,2015,Identification and analysis of icCu/Zn-SOD,Mn-SOD and ecCu/Zn-SOD in superoxide dismutase multigene family of Pseudosciaena crocea,Fish Shellfish Immunol.,43(2):491-501.
Malhotra J.D.,and Kaufman R.J.,2007,Endoplasmic reticulum stress and oxidative stress:a vicious cycle or a double-edged sword?Antioxid.Redox Signal.,9(12):2277-2293.
Mccord J.M.,and Fridovich I.,1969,Superoxide dismutase.An enzymic function for erythrocuprein(hemocuprein),Journal of Biological Chemistry,244(22):6049-6055.
Nakamura H.,and Nakamura K.J.,1997,Redox regulation of cellular activation,Annual Review of Immunology,15(2):351-369.
Pérez-Sánchez J.,Borrel M.,Bermejo-Nogales A.,Benedito-Palos L.,Saera-Vila A.,Calduch-Giner J.A.,and Kaushik S.,2013,Dietary oils mediate cortisol kinetics and the hepatic m RNA expression profile of stress-responsive genes in gilthead sea bream(Sparus aurata) exposed to crowding stress,Implications on energy homeostasis and stress susceptibility,Comparative Biochemistry&Physiology Part D Genomics&Proteomics,8(2):123-130.
Reyes-Becerril M.,Maldonado-García M.,Guluarte C.,León-Gallo A.,Rosales-Mendoza S.,Ascencio F.,Hirono I.,and Angulo C.,2016,Evaluation of ToxA and vibrio parahaemolyticus lysate on humoral immune respons e and immune-related genes in Pacific red snapper,Fish Shellfish Immunol.,56:310-321.
Wang Q.,Yuan Z.,Wu H.,Liu F.,and Zhao J.,2013,Molecular characterization of a manganese superoxide dismutase and copper/zinc superoxide dismutase from the mussel Mytilus galloprovincialis,Fish Shellfish Immunol.,34(5):1345-1351.
Zhang Z.W.,Li Z.,Liang H.W.,Li L.,Luo X.Z.,and Zou G.W.,2011,Molecular cloning and differential expression patterns of copper/zinc superoxide dismutase and manganese superoxide dismutase in Hypophthalmichthys molitrix,Fish Shellfish Immunol.,30(2):473-479.
Zhang H.Y.,Yu X.D.,Wang P.P.,Jia X.Q.,Li X.R.,Cao Q.Q.,Yin S.W.,Chen S.Q.,and Zhou G.Q.,2017,Molecular cloningand sequence analysis of Mn-SOD gene from Odontobutis potamophila,Yuye Kexue Jinzhan(Progress in Fishery Sciences),38(4):34-43.(张宏叶,于兴达,王佩佩,贾秀琪,李欣茹,曹全全,尹绍武,陈树桥,周国勤,2017,河川沙塘鳢(Odontobutis potamophila) Mn-SOD基因的克隆及表达分析,渔业科学进展,38(4):34-43.)
基本信息:
DOI：10.13417/j.gab.040.002020
中图分类号:S917.4
引用信息:
[1]朱祎,刘伊凡,王怡,等.布氏鲳鲹超氧化物歧化酶基因的序列分析及组织分布[J].基因组学与应用生物学,2021,40(Z1):2020-2028.DOI:10.13417/j.gab.040.002020.
基金信息:
海南省重点研发计划课题(ZDYF2017038); 南海海洋资源利用国家重点实验室开放课题(2018001); 国家“863”计划项目(2012AA10407)共同资助
2019-03-07
2019-03-07
2019-03-07