中华绒螯蟹类浮舰蛋白基因Esflol的克隆和组织表达分析

Cloning and expression analysis of Esflol gene in Eriocheir sinensis

为了解甲壳动物高血糖激素如何调节中华绒螯蟹体内的血糖水平,实验前期进行了转录组和表达谱分析,最终选取Esflol作为研究对象。根据转录组数据提示,结合RACE方法首次从中华绒螯蟹肝胰腺组织中克隆得到Esflol基因并进行了序列及组织表达分析。结果显示Esflol基因开放阅读框(ORF)1 281 bp,编码426个氨基酸,具有典型的SPFH超家族和PHB结构域,不含信号肽序列。多重序列比对分析表明,Esflol氨基酸序列与中华绒螯蟹flotillin-1相似度最高,达到57%。荧光定量PCR结果显示成熟中华绒螯蟹Esflol基因在肝胰腺组织中的表达量最高,其次为肌肉,另外在心脏、鳃、肠道、胸神经节和胃中均有一定量表达,在脑中表达量较低。上述实验结果为下一步继续研究中华绒螯蟹糖代谢的生理机制奠定了基础,也为其他养殖类甲壳动物糖类代谢的研究提供了重要参考。

The crustacean hyperglycemic hormone（ CHH） family has been found only in arthropods currently,and plays significant roles in regulation of glucose levels,molting,stress response and a variety of physiological metabolic activities. We selected Esflol（ Eriocheir sinensis flotillin-1 like,Esflol） as our object of study afer transcriptome analysis and gene expression profiling,to understand the importance of this family —crustacean hyperglycemic hormone（ CHH）,howto regulate glucose levels of the E. sinensis.According to the prompting of transcriptiome,a cDNA sequence encoding Esflol was cloned from hepatopancreas of E. sinensis by RACEs,and the results of tissue expression analysis were also obained. The results showthat the open reading frame of Esflol consists of 1281 bp,encoding 426 amino acids without signal peptide. Typical domains of SPFH super family and PHB domain. Multi-sequence alignments analysis showed Esflol shared the higest identity with E. sinensis flotillin-1（ 57%）. Tissue distribution of Esflol was detected by quantitative real-time PCR,although strong expression of Esflol was detected in hepatopancreas and muscles,and its expression could also be found in heart,gill,gut,ganglion and stomach,and in lower expression levels of brain. This study lays the foundation for sugar metabolic physiological mechanism research of E. sinensis,and also provides a reference for sugar metabolic physiological mechanism research of other aquaculture crustaceans.