大鲵泛素结合酶cgsUBE2L3基因克隆及其在免疫应答中的表达分析  

Cloning and Expression of Ubiquitin-Conjugating Enzyme cgsUBE2L3 Gene in Chinese Giant Salamander (Andrias davidianus) During the Immune Response

作　　者：刘榴[1] 石君玲 尤修玲 廖志勇[1] LIU Lin SHI Jun-ling YOU Xin-ling LIAO Zhi-yong(College of Life and Environmental Science, Wenzhou University, Zhejiang Provincial Key Laboratory for Water Environment and Marine Biological Resources Protection, Wenzhou, Zhejiang 32503)

机构地区：[1]温州大学生命与环境科学学院,浙江省水环境与海洋生物资源保护重点实验室,浙江温州325035

出　　处：《安徽农业科学》2017年第22期93-97,154,共6页Journal of Anhui Agricultural Sciences

基　　金：国家重点基础研究计划(973计划)子课题(2013CB835301)

摘　　要：[目的]研究泛素结合酶UBE2L3基因在大鲵免疫应答过程中的作用。[方法]克隆大鲵泛素结合酶基因(cgsUBE2L3),进行相关生物信息学分析,并通过qRT-PCR对其组织分布及免疫刺激下的表达谱进行分析。[结果]成功克隆了cgsUBE2L3基因,该基因全长包含一个462 bp的开放阅读框,编码154个氨基酸,预计蛋白分子量为21.2 ku;cgsUBE2L3氨基酸序列保守性很高,与非洲爪蟾的同源性高达97.4%;此外,qRT-PCR结果显示,cgsUBE2L3基因在大鲵组织中广泛分布,且被LPS感染的大鲵多个免疫组织中cgsUBE2L3的mRNA水平均发生了明显变化。[结论]cgsUBE2L3基因具有高度保守的结构特征,其组织分布具有特异性且参与大鲵抵抗LPS刺激的免疫应答。[ Objective] To study the role of ubiquitin-conjugating enzyme cgsUBE＇21-3 gene during the immune response in Chinese giant salamander （Andrias davidianus）. [ Method] To clone ubiquitin-eonjugating enzyme gene from Chinese giant salamander （cgsUBE＇2L3） and analyzed its amino acids by bioinformatics software, the tissue distribution and expression profile with the stimulation of lipopolysaceharide （LPS） were detected by qRT-PCR. [ Result] Successfully, we cloned cgsUBE2L3 gene. The full-length cDNA ofcgsUBE2L3 contained an open reading frame of 462 bp, encoding a protein of 154 amino acids with a putative weight of 21.2 ku. The cgsUBE2L3 had a higldy conserved amino acid sequence and shared the highest identity of 97.4% with Mrican clawed frog. Quantitative real time PCR results showed that the cgsUBE2L3 gene was widely distributed in the Chinese giant salamander tissues and the mRNA levels ofcgsUBE2L3 were significantly changed in the immu-ne-associated tissues of LPS-infeeted Chinese giant salamander. [ Conclusion] The cgsUBE2L3 gene has highly conserved structural feature and tissue-specific of mRNA expression. Moreover, it participates in the immune response of Chinese giant salamander to resist LPS stimulation.

关 键 词：大鲵 UBE2L3 序列分析 免疫应答 

分 类 号：S966.6[农业科学—水产养殖]