花鳗鲡AQP3基因的分子特征及其应对盐度变化的表达规律

发布时间：2018-08-04 18:17

【摘要】：为了探究花鳗鲡(Anguila marmorata)AQP3基因在不同盐度下的表达规律,利用RACE技术克隆了花鳗鲡AQP3基因c DNA全长序列,并进行了生物信息学分析;通过实时荧光定量PCR(RT-q PCR),检测了花鳗鲡AQP3在9个组织中的分布及盐度胁迫后鳃、肾、肠组织在0、6、12、24 h和3、5、10 d的表达情况。结果显示:花鳗鲡AQP3基因的c DNA全长为1299 bp,开放阅读框(ORF)为891 bp,5’非编码区(UTR)和3’非编码区(UTR)分别为60 bp和348 bp,共编码296个氨基酸,具有6段跨膜结构域及2个NPA(天冬氨酸-脯氨酸-丙氨酸)保守结构域。氨基酸多重序列比对结果显示,花鳗鲡AQP3基因与其同属的欧洲鳗鲡(Anguilla anguilla)、日本鳗鲡(Anguilla japonica)同源性分别高达97.0%和96.0%。RT-q PCR检测发现,AQP3基因在鳃组织中表达量最高,而在脾脏中表达量最低;在咸淡水(盐度10)和海水(盐度25)组中,鳃组织的AQP3基因表达水平呈下降趋势,且在各时段均显著下调;肾组织的AQP3表达水平在咸淡水(除24 h外)和海水(除6 h外)组中也显著下调;肠组织的AQP3表达水平在咸淡水组中显著上调,而在海水组中则显著下调。研究表明,花鳗鲡AQP3的mRNA表达量在应对不同盐度环境时呈现不同的表达变化模式,这将为深入揭示花鳗鲡适应盐度变化的分子调控机理奠定理论基础。
[Abstract]:In order to investigate the expression of (Anguila marmorata) AQP3 gene in Anguilla japonica under different salinity, the full-length sequence of c DNA of AQP3 gene of Anguilla Anguilla japonica was cloned by RACE technique and analyzed by bioinformatics. The distribution of AQP3 in 9 tissues and the expression of AQP3 in the Gill, kidney and intestine of Anguilla japonica were detected by real-time fluorescence quantitative PCR (RT-q PCR),). The results showed that the total length of c DNA of the AQP3 gene of Anguilla japonica was 1299 BP, the open reading frame (ORF) was 891 BP (UTR) and the 3'noncoding region (UTR) was 60 BP and 348 BP, respectively, encoding 296 amino acids. There are six transmembrane domains and two NPA conserved domains. The results of amino acid multiple sequence alignment showed that the homology of the AQP3 gene of Anguilla japonica was 97.0% with that of the European eel (Anguilla anguilla), of the same genus, and the highest expression of (Anguilla japonica) gene was found in the Gill tissue of Japanese Anguilla japonica and the lowest in the spleen by 96.0%.RT-q PCR test. In saltwater (salinity 10) and seawater (salinity 25) groups, the expression of AQP3 gene in Gill tissue showed a decreasing trend, and was significantly down-regulated at each stage. The expression of AQP3 in renal tissues was also significantly down-regulated in saline and fresh water (except 24 h) and seawater (except 6 h) groups, while the expression of AQP3 in intestinal tissues was significantly up-regulated in saline and fresh water groups, but significantly down-regulated in seawater groups. The results showed that the expression of mRNA in AQP3 of Anguilla japonica showed different expression patterns in different salinity environments, which would lay a theoretical foundation for further understanding the molecular regulation mechanism of adaptation to salinity change in Anguilla Anguilla japonica.
【作者单位】：南京师范大学生命科学学院江苏省生物多样性与生物技术重点实验室;
【基金】：江苏省自然科学基金项目(BK20141450)
【分类号】：S917.4

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