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.
Abstract
[Objective] To study the role of ubiquitin-conjugating enzyme cgsUBE2L3 gene during the immune response in Chinese giant salamander (Andrias davidianus).[Method] To clone ubiquitin-conjugating enzyme gene from Chinese giant salamander (cgsUBE2L3) and analyzed its amino acids by bioinformatics software,the tissue distribution and expression profile with the stimulation of lipopolysaccharide (LPS) were detected by qRT-PCR.[Result] Successfully,we cloned cgsUBE2L3 gene.The full-length cDNA of cgsUBE2L3 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 highly conserved amino acid sequence and shared the highest identity of 97.4% with African 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 of cgsUBE2L3 were significantly changed in the immu-ne-associated tissues of LPS-infected 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.
基金项目
国家重点基础研究计划(973计划)子课题(2013CB835301)
国家科技期刊平台
NSTL
维普
万方数据