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斜带石斑鱼MyD88基因的克隆与表达
Cloning and Expression of MyD88 Gene in Orange-spotted Grouper Epinephelus coioides

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韦友传 1   高谦 2   昌明先 2   罗廷荣 3 *  
文摘 本研究运用RACE-PCR技术获得斜带石斑鱼(Epinephelus coioides)髓样分化因子88(myeloid differentiation factor 88,MyD88)基因,并对该基因进行生物信息学和表达模式分析。研究结果表明1795bp的cDNA全长序列,包括ORF 870 bp、5’UTR 243 bp和3’UTR 682 bp,3’UTR存在1个多聚腺苷酸加尾信号(AATAAA)和两个mRNA不稳定基序(ATTTA)。SMART软件预测该蛋白N端和C端分别存在死亡结构域和TIR结构域(Toll/IL-1 receptor homology domain,TIR);与其它脊椎动物MyD88的序列同一性达57.1%～78.7%;用NJ法构建的系统进化树中,斜带石斑鱼MyD88和其它已报导的鱼类MyD88聚为一枝。qPCR检测结果显示MyD88基因mRNA主要表达于肝脏、脾脏、头肾和胸腺等组织。本研究为进一步探讨MyD88在斜带石斑鱼TLR信号传导中的作用奠定基础。
其他语种文摘 The full cDNA sequence of myeloid differentiation factor 88(MyD88)in the orange-spotted grouper Epinephelus coioides,designated as EcMyD88,has been isolated by using RACE-PCR method.The EcMyD88 consisted of 1 795 bp,including an 870 bp open reading frame(ORF),a 243 bp 5’ UTR and a 682 bp 3’ UTR.The 3’ UTR contained a polyadenylation-tailed signal(AATAAA)and 2 mRNA instable motifs(ATTTA).The EcMyD88 protein comprised a death domain on N terminal and a TIR domain on C terminal by using SMART program.Putative amino acid sequence of EcMyD88 shared 57.1%～78.7% identity with its counterparts from other vertebrates.Phylogenetic tree was constructed by using neighbor-joining method which revealed that EcMyD88 was clustered with MyD88 from other teleost fish reported previously.The transcription of EcMyD88 was examined by real-time quantitative PCR,and its mRNA was mainly expressed in liver,spleen,thymus and head kidney.This study will lay a foundation for further exploring the role of MyD88 in the TLR signalling in Epinephelus coioides.
来源 基因组学与应用生物学 ,2011,30(3):288-295 【扩展库】
关键词 斜带石斑鱼 ; 表达 ; 髓样分化因子88(MyD88)
地址

1. 广西大学亚热带生物资源保护利用重点实验室, 淡水生态与生物技术国家重点实验室, 南宁, 530004  

2. 中国科学院水生生物研究所, 淡水生态与生物技术国家重点实验室, 武汉, 430072  

3. 广西大学亚热带生物资源保护利用重点实验室, 南宁, 530004

语种 中文
文献类型 研究性论文
ISSN 1674-568X
学科 分子生物学;水产、渔业
基金 国家自然科学基金 ;  广东省人民政府自然科学联基金
文献收藏号 CSCD:4259186

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引证文献 7 篇

1 韦友传 草鱼hepcidin基因cDNA克隆、序列分析与表达 上海海洋大学学报,2012,21(4):495-501
被引 0 次

2 韦友传 斜带石斑鱼MyD88基因的原核表达及蛋白纯化 水生生物学报,2014,38(1):200-202
被引 3 次

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