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为研究水通道蛋白11基因(AQP11)在中华绒螯蟹(Eriocheir sinensis)生长蜕壳过程中的功能作用,采用RACE技术克隆获得中华绒螯蟹水通道蛋白11基因cDNA全长序列。该序列总长为1 746 bp,5'端和3'端非编码区分别为463 bp和476 bp,开放阅读框为807 bp,推测编码268个氨基酸,预测分子量29.46 kDa,理论等电点为5.38。生物学信息分析表明,AQP11含有4个跨膜区(第62～84,第159～181,第194～216,第231～250)和2个NPV单元,属于稳定蛋白;同源性和进化树分析表明,中华绒螯蟹AQP11氨基酸序列与凡纳滨对虾(Litopenaeus vannamei)的同源性最高(82.0%),与凡纳滨对虾的聚为一支,与甲壳动物的亲缘关系最近。实时荧光定量PCR (RT-qPCR)的检测显示,AQP11基因在中华绒螯蟹各组织中均有表达,其中在肠道中表达量最高,其次是脑、肌肉和胸神经节,在肝胰腺、鳃和血中表达量最低。研究发现,AQP11基因在中华绒螯蟹肠道中的表达呈现,在蜕壳间期(C期)和蜕壳前期(D期)过程中表达量均较低,在蜕壳期(E期)表达量开始上升,蜕壳后期(AB期)表达量不变。AQP11基因在肌肉中的表达呈现,蜕壳间期(C期)表达量低,蜕壳前期(D期)表达量开始上升,蜕壳期(E期)达到峰值,随后到蜕壳后期(AB期)下降。研究结果表明,中华绒螯蟹AQP11基因在其蜕壳过程中发挥着重要的作用。
Abstract：In order to study the functional role of aquaporin 11 gene( AQP11) in the process of growth and molting of Eriocheir sinensis, the full-length c DNA sequence of aquaporin 11 gene of E. sinensis was cloned by RACE technology. The full-length of AQP11 c DNA was 1 746 bp, with a 463 bp 5'-untranslated region(UTR), a 476 bp3'-UTR, and an open reading frame(ORF) of 807 bp which encoded a 268 amino acid polypeptide, the predicted molecular weight was 29.46 kDa, and the theoretical isoelectric point was 5.38. Bioinformatics software analysis revealed that AQP11 gene contained 4 transmembrane domains(62～84, 159～181, 194～216, 231～250), 2 NPV structural units, being part of stable proteins. Homologous analysis showed that AQP11 amino acid sequence of E.sinensis had the highest homology to Litopenaeus vannamei(82.0%), clustered with L. vannamei, and the closest relationship with crustaceans. Real-time quantitative(RT-qPCR) showed that AQP11 gene could be detected in all tested tissues of Eriocheir sinensis, with the highest expression level in intestine, followed by the brain, muscle and thoracic ganglion, and the lowest expression level in the hepatopancreas, gills and blood. In the intestinal, the expression of AQP11 was significantly lower at the intermolt stage(stages C) and pre-molt stage(stage D), significantly enhancing and reaching the maximal level at the ecdysis(stage E). On the other hand, the expression of AQP11 gene in muscle showed low expression in the inter-molt stage(stage C), and increased gradually at the pre-molt stage(stage D), significantly enhancing and reaching the maximal level at the ecdysis stage(E stage), and then reaching the postmolt stage(stage A and B) decline. The results indicated that the AQP11 gene played an important role in the molting process of E. sinensis.
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基本信息:
DOI：10.13417/j.gab.040.001497
中图分类号:S917.4
引用信息:
[1]张龙,庞杨洋,杨航,等.中华绒螯蟹水通道蛋白11基因克隆及表达分析[J].基因组学与应用生物学,2021,40(04):1497-1504.DOI:10.13417/j.gab.040.001497.
基金信息:
国家重点研发计划项目(2018YFD0900603); 现代农业产业技术体系专项资金项目(CARS-48); 上海市科委工程技术中心能力提升项目(19DZ2284300)共同资助
2021-04-25
2021-04-25