斑节对虾Chitinase-2基因的克隆及其在蜕皮和幼体发育过程中的表达分析

发布时间：2018-03-20 16:18

本文选题：斑节对虾　切入点：几丁质酶　出处：《南方水产科学》2017年04期 　论文类型：期刊论文

【摘要】：研究以斑节对虾(Penaeus monodon)转录组获得的几丁质酶基因(Pm Chi)片段,运用RACE技术克隆了PmChi基因c DNA全长,命名为PmChi-2。PmChi-2基因c DNA全长2 050 bp,其中5'-非编码区(5'-UTR)144 bp、3'-UTR 319 bp和开放阅读框(ORF)1 587 bp,编码528个氨基酸。生物信息学分析显示,PmChi-2与其他甲壳动物Chi-2的相似性为78%~97%。实时荧光定量PCR结果显示,PmChi-2在蜕皮前期(D期)表皮中表达水平最高,在胃、鳃、腹神经节和眼柄中表达水平依次降低,在其他组织(肝胰腺、肠、肌肉、心脏)中几乎不表达。不同蜕皮阶段,PmChi-2在鳃、胃和表皮3种组织中的表达变化模式基本一致,均在蜕皮期(E期)表达量最低,而最高表达量在鳃中出现在蜕皮后期(A期),在胃和表皮中为D期。幼体不同发育阶段分析揭示PmChi-2 mRNA在幼体发育阶段的无节幼体到糠虾幼体第二期表达量维持在一个低水平,在糠虾幼体第三期PmChi-2 mRNA表达水平显著升高,在仔虾期又显著下降,推测PmChi-2 mRNA可能与斑节对虾幼体发育密切相关。研究结果表明PmChi-2基因可能在斑节对虾蜕皮以及幼体的变态发育中发挥重要作用,为深入研究斑节对虾几丁质酶发育调控提供了重要信息。
[Abstract]:The fragment of chitinase gene Pm Chiwas obtained from Penaeus monodon transcriptome, and the full length of PmChi gene c DNA was cloned by RACE technique. The c DNA of PmChi-2.PmChi-2 gene was named as 2 050 BP, of which the 5 PmChi-2.PmChi-2 non-coding region was 144bpt3 + -UTR319 BP and the open reading frame ORFN 1 587 BP, encoding 528 amino acids. Bioinformatics analysis showed that the similarity between PmChi-2 and other crustacean Chi-2 was 78797. The real-time fluorescence was used to determine the similarity between PmChi-2 and other crustaceans. The results of bioinformatics analysis showed that the similarity between PmChi-2 and other crustaceans was 7897. The results of quantitative PCR showed that PmChi-2 expression was the highest in the epidermis of preecdylastic stage D). The expression level in stomach, Gill, ventral ganglion and eye stalk decreased in turn, but hardly in other tissues (hepatopancreas, intestine, muscle, heart). The expression patterns of gastric and epidermal tissues were basically the same, and the expression levels were the lowest in E stage of molting. The highest expression level was found in the Gill at the late stage of decidua and D in the stomach and epidermis. The analysis of the different stages of larval development revealed that the expression of PmChi-2 mRNA was maintained at a low level in the second stage of larval stage from the stage of larval development to that of the second stage of larval stage of furfural shrimp. The expression of PmChi-2 mRNA increased significantly in the third stage of the larvae, and decreased in the larval stage. It is speculated that PmChi-2 mRNA may be closely related to juvenile development of Penaeus vannamei. The results suggest that the PmChi-2 gene may play an important role in the ecdysis and metamorphosis of Penaeus mongolicus. It provides important information for the further study of chitinase regulation of Penaeus Monodon.
【作者单位】：中国水产科学研究院南海水产研究所农业部南海渔业资源开发利用重点实验室;上海海洋大学水产与生命学院;
【基金】：现代农业产业技术体系建设专项资金(CARS-47) 广东省省级科技计划项目(2014B020202003) 国家高技术研究发展计划(863计划)项目(2012AA10A409) 广东省海洋与渔业科技推广专项(A201501A06) 深圳市生物产业发展专项资金现代农业生物产业推广扶持计划项目(NYSW201400331010053) 海南省应用技术研法与示范推广专项(ZDXM2015056)
【分类号】：S917.4

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