瘤背石磺Egr-1基因的克隆、分析及低频声音刺激对其表达的影响  

Cloning and Expression Analysis of the Egr-1 Gene of Onchidium reevesii and Low Frequence Sound Stimulaton on Gene Expression

作　　者：吴容宇 梁威 土志涵[1,2,3] 杨铁柱 沈和定[1,2,3] Wu Rongyu;Liang Wei;Tu Zhihan;Yang Tiezhu;Shen Heding(Key Lab of Ministry of Education for Exploration and Utilization of Aquatic Genetic Resources,Shanghai Ocean University,Shanghai,201306;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai,201306;Key Laboratory of Marine Animal Taxonomy and Evolution,Shanghai Ocean University,Shanghai,201306)

机构地区：[1]上海海洋大学,水产种质资源发掘与利用教育部重点实验室,上海201306 [2]上海海洋大学,水产科学国家级实验教学示范中心,上海201306 [3]上海海洋大学,海洋动物系统分类与进化上海高校重点实验室,上海201306

出　　处：《基因组学与应用生物学》2021年第2期622-630,共9页Genomics and Applied Biology

基　　金：国家自然科学基金项目(41276157)资助。

摘　　要：早期生长反应因子1(Egr-1)基因又名为Zif268、NGFI-A、Krox-24、Tis8、ZENK,分布于脊椎动物的神经细胞中,参与识别功能相关的神经活动。本研究通过c DNA末端快速扩增法(RACE)获得瘤背石磺(Onchidium reevesii)Egr-1基因全长序列,进行蛋白质特征分析、氨基酸序列预测以及Egr-1基因的组织差异表达,并检测了不同低频声音(0~200 Hz)刺激0~24 h后Egr-1基因表达量的变化。结果显示:瘤背石磺Egr-1基因c DNA序列全长2556 bp,含134 bp的5’端非编码区、709 bp的3’端非编码区和1713 bp的开放阅读框,编码570个氨基酸。瘤背石磺Egr-1蛋白分子质量为61.31 kD,理论等电点为6.14,平均疏水指数-0.491,不含信号肽;二级结构和三级结构预测显示有琢螺旋和茁折叠。瘤背石磺Egr-1基因在其腹足、背部皮肤、神经节、肝胰腺、两性腺中均有表达,在神经节和两性腺中高表达,背部皮肤和肝胰腺中组织表达量最低(P<0.05)。相比对照组,受到低频声音刺激0.5 h,后瘤背石磺神经节中Egr-1基因的表达量显著升高,且不同频率声音间表达量差异显著(P<0.05),推测Egr-1基因参与了瘤背石磺识别低频声音功能相关的神经活动,所得结果为进一步研究瘤背石磺识别处理声音信息的分子机制提供重要的基础数据。The early growth response 1(Egr-1)gene,also known as Zif268,NGFI-A,Krox-24,Tis8,ZENK,is distributed in the vertebrate nerve cells and participates in recognition functionally related neural activities.In this study,the full-length sequence of Egr-1 gene in Onchidium reevesii was cloned by using rapid amplification of cDNA end(RACE)method,and the amino acid sequence and protein characteristics were also evaluated.Furthermore,the gene expression of Egr-1 were studied before and after 0~24 h stimulation of different low frequencies sound(0~200 Hz).The results showed that the full-length cDNA sequence of Egr-1 gene was 2256 bp,which contained 134 bp 5’non-coding region,709 bp 3’non-coding region,1713 bp open reading frame,and encoding 570 amino acids.The molecular mass of Egr-1 protein was 61.31 kD,the theoretical isoelectric point was 6.14,the average hydrophobic index was-0.491,and no signal peptide was contained.Its secondary and tertiary structures included alpha helix and beta strand.The gene expressed of Egr-1 in O.reevesii was detected in pleopod dorsal skin,ganglion,hepatopancreas and hermaphroditic organ,and it was significantly higher in ganglion and hermaphroditic organ,but in dorsal skin and hepatopancreas was the lowest(P<0.05).AfterO.reevesii receives0.5 h of low frequence sound stimulation,the gene expression of Egr-1 in ganglion was significantly higher than the control group,and the expression levels of different frequencies sounds were significantly different at this time(P<0.05).It is speculated thatEgr-1 genes involved in the neural activities related to low frequencies sound perception of O.reevesii,and the results provide a basis for further research on the molecular mechanism of sound information perception in O.reevesii.

关 键 词：瘤背石磺 EGR-1 低频声音感知 基因克隆 

分 类 号：S917.4[农业科学—水产科学]