Cloning, genomic structure and expression analysis of ubc9 in the course of development in the half-smooth tongue sole (Cynoglossus semilaevis)
Cloning, genomic structure and expression analysis of ubc9 in the course of development in the half-smooth tongue sole (Cynoglossus semilaevis)
复制标题
半滑舌鳎（Cynoglossus semilaevis）发育过程中ubc9的克隆、基因组结构及表达分析
DOI：
10.1016/j.cbpb.2013.03.007
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发表时间：
2013-07-01
影响因子：
2.2
通讯作者：
Chen, Songlin
中科院分区：
文献类型：
--
作者：
Hu, Qiaomu;Chen, Songlin
小泛素样修饰物（SUMO）通路是真核生物中一个重要的生物学过程，Ubc9是SUMO通路中一种重要的E2结合酶（UBE2），在多种生物的细胞分化、发育以及性别调控中发挥关键作用。然而，Ubc9与鱼类性别调控和发育之间的关系仍不清楚。为了阐明Ubc9对性别调控和发育的影响，从半滑舌鳎（Cynoglossus semilaevis）中克隆了其cDNA全长和基因组序列。实时定量RT - PCR表明，ubc9在不同组织中普遍表达，并且在胚胎和性腺发育的不同阶段表达水平存在差异。此外，在经过温度处理的雌性个体中其表达水平显著高于正常雌性和雄性。而且，构建了PET - 32 - Ubc9质粒，并在大肠杆菌中表达了重组蛋白。注射Ubc9蛋白后，卵泡抑素基因表达最初上调，而FSE基因（cyp19a1a、ctnnb1、foxl2）最初下调，在96小时之前最终恢复到正常水平。综上所述，结果表明Ubc9参与胚胎发生、配子发生和性别调控，并对基因表达产生影响。（c）2013爱思唯尔公司。保留所有权利。
The small ubiquitin-like modifier (SUMO) pathway is an essential biological process in eukaryote, and Ubc9 is an important E2 conjugating enzyme (UBE2) for SUMO pathway and plays a critical role in cellular differentiation, development and sex modification in various species. However, the relationship between Ubc9 and sex modification and development in fish remains elusive. To elucidate the impact of Ubc9 on sex modification and development, the full length of the cDNA and genomic sequence was cloned from half-smooth tongue sole, Cynoglossus semilaevis. Real-time quantitative RT-PCR demonstrated that ubc9 was ubiquitously expressed in different tissues, and the expression levels varied in the different stages of embryonic and gonadal development. In addition, the expression level was significantly higher in the temperature-treated females than the normal females and males. Moreover, the PET-32-Ubc9 plasmid was constructed and the recombinant protein was expressed in Escherichia coli. Follistatin gene expression was initially up-regulated and FSE genes (cyp19a1a, ctnnb1, foxl2) were initially down-regulated after the injection of Ubc9 protein, prior to 96 h eventually recovered to normal levels. Taken together, the results show that Ubc9 is involved in embryogenesis, gametogenesis and sex modification, and exerts an effect on gene expression. (c) 2013 Elsevier Inc. All rights reserved.