尼罗罗非鱼Cullin-1基因克隆及表达Cloning and Tissue Expression Analysis of the Cullin-1 in Nile Tilapia (Oreochromis niloticus)
刘鑫潮,黄瑜,简纪常
摘要(Abstract):
【目的】探究尼罗罗非鱼(Oreochromis niloticus)Cullin-1(CUL1)基因在尼罗罗非鱼抵抗病原细菌过程中的作用。【方法】从尼罗罗非鱼脾脏组织中克隆Cullin-1基因(记为On-CUL1),对其进行生物信息学分析,应用实时荧光定量PCR技术分析On-CUL1在健康鱼胸腺、鳃、肝、脾、肠道、头肾、脑、肌肉、皮肤、血液组织,以及经无乳链球菌刺激后病鱼脾、肠道、头肾中的表达。【结果与结论】On-CUL1(GenBank登录号XM_003459470.5)编码区为2 331 bp,编码776个氨基酸,理论分子质量为89 ku,理论等电点为8.16。氨基酸序列分析显示,On-CUL1有1个高度保守的泛素样蛋白结构域CUL1 Nedd8(ubiquitin-like domain),N′端有一个的信号肽序列。系统进化树分析表明,On-CUL1序列与伯氏拟丽鱼(Haplochromisburtoni)CUL1相似性最高,为98.7%。q RT-PCR显示CUL1在健康尼罗罗非鱼各组织中均有表达,其中在头肾中表达量最高,其次是胸腺、脑、皮肤,在肠道中表达量最低。经无乳链球菌刺激后,On-CUL1表达量在脾脏和肠道中均显著上调,头肾、脾脏和肠道均在48h表达量最大。CUL1参与尼罗罗非鱼抵抗无乳链球菌过程中的免疫应答。
关键词(KeyWords): 尼罗罗非鱼;CUL1;基因表达;免疫应答;无乳链球菌
基金项目(Foundation): 国家自然科学基金(31572651);; 高校重大科研成果培育计划类项目(GDOU2013050309);; 广东省普通高校青年创新人才项目(230419083)
作者(Author): 刘鑫潮,黄瑜,简纪常
参考文献(References):
- [1]黄家庆,徐田祥.小体积高密度网箱养殖尼罗罗非鱼技术[J].齐鲁渔业,2004,21(7):17-19.
- [2]张新艳,樊海平,钟全福,等.罗非鱼无乳链球菌的分离、鉴定及致病性研究[J].水产学报,2008,32(5):772-779.
- [3]黄锦炉.罗非鱼无乳链球菌病病原学、病理学及cps E基因的原核表达研究[D].雅安:四川农业大学,2012.
- [4]谢彩霞,汪志文,鲁义善,等.尼罗罗非鱼pik3r1基因的克隆和m RNA表达分析[J].广东海洋大学学报,2020,40(1):8-14.
- [5]周蕊,余泽华.泛素化途径与细胞周期的关系[J].生命科学,2003,15(3):147-150.
- [6]ZHOU Y H,XIA J Z,XU W H,et al.Cullin-1 promotes cell proliferation in human breast cancer and is related to diabetes[J].International Journal of Biological Markers,2016,31(4):375-381.
- [7]雍红梅.Cullin1基因在乳腺癌发生发展中功能及分子机制研究[D].徐州:徐州医学院,2012.
- [8]WANG W,LIU Z H,QU P,et al.Knockdown of regulator of cullins-1(ROC1)expression induces bladder cancer cell cycle arrest at the G2 phase and senescence[J].PLo SOne,2013,8(5):1-11.
- [9]汪志文,张海艳,黄瑜,等.尼罗罗非鱼CD80基因的分子克隆与m RNA表达分析[J].基因组学与应用生物学,2017,36(1):190-200.
- [10]HANSEN J D,KAATTARI S L.The recombination activating gene 1(RAG1)of rainbow trout(Oncorhynchus mykiss):cloning,expression,and phylogenetic analysis[J].Immunogenetics,1995,42(3):188-195.
- [11]HANSEN J D,KAATTARI S L.The recombination activating gene 2(RAG2)of the rainbow trout Oncorhynchus mykiss[J].Immunogenetics,1996,44(3):203-211.
- [12]HANSEN J D,STRASSBURGER P,DU PASQUIER L.Conservation of a master hematopoietic switch gene during vertebrate evolution:isolation and characterization of Ikaros from teleost and amphibian species[J].European Journal of Immunology,1997,27(11):3049-3058.
- [13]GAN Z,WANG B,ZHOU W,et al.Molecular and functional characterization of CD59 from Nile tilapia(Oreochromis niloticus)involved in the immune response to Streptococcus agalactiae[J].Fish&Shellfish Immunology,2015,44(1):50-59.
- [14]CESTA M F.Normal structure,function,and histology of the spleen[J].Toxicologic Pathology,2006,34(5):455-465.
- [15]SCHUCHAT A.Epidemiology of group B streptococcal disease in the United States:shifting paradigms[J].Clinical Microbiology Reviews,1998,11(3):497-513.
- [16]ROBINSON J A,MEYER F P.Streptococcal fish pathogen[J].Journal of Bacteriology,1966,92(2):512-513.
- [17]KOZONO Y,ABE R,KOZONO H,et al.Cross-linking CD21/CD35 or CD19 increases both B7-1 and B7-2expression on murine splenic B cells[J].Journal of Immunology,1998,160(4):1565-1572.
- [18]罗晓春,谢明权,黄玮,等.鱼类粘膜免疫研究进展[J].水产学报,2005,29(3):411-416.
- [19]MERINO-CONTRERAS M L,GUZMAN-MURILLOM A,RUIZ-BUSTOS E,et al.Mucosal immune response of spotted sand bass Paralabrax maculatofasciatus(Steindachner,1868)orally immunised with an extracellular lectin of Aeromonas veronii[J].Fish&Shellfish Immunology,2001,11(2):115-126.
- [20]唐玫,马广智,徐军.鱼类免疫学研究进展[J].免疫学杂志,2002,18(z1):112-116.
- [21]樊博琳,黎源,汪志文,等.尼罗罗非鱼CD247基因克隆及其组织表达分析[J].广东海洋大学学报,2018,38(6):6-12.