摘要
超氧化物歧化酶(SOD)是通过歧化反应催化超氧化物转化为O_(2)和H_(2)O_(2)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。
Superoxide dismutases(SOD)are some enzymes that catalyze superoxide to oxygen(O_(2))and hydrogen peroxide(H_(2)O_(2))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.
作者
朱祎
刘伊凡
王怡
叶恒振
符丹凤
骆剑
Zhu Yi;Liu Yifan;WangYi;Ye Hengzhen;Fu Danfeng;Luo Jian(Key Laboratory of for Ministry of Education for Tropical Biological Resources,State Key Laboratory of Marine Resource Utilization in South China Sea,Ocean College of Hainan University,Haikou,570228)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2021年第5期2020-2028,共9页
Genomics and Applied Biology
基金
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