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中华绒螯蟹C型凝集素基因组和启动子的克隆与分析

作者:时间:2018-05-04点击数:

PDF全文下载:  201803009.pdf

文章编号: 16726987201803005307 DOI 10.16351/j.16726987.2018.03.009

 

李凤梅, 祝倩倩

(青岛科技大学 海洋科学与生物工程学院,山东 青岛 266042

 

摘要: 为探究中华绒螯蟹(Eriocheir sinensis)凝集素(Lectin)基因启动子在免疫应答中的转录调控机制,本研究从中华绒螯蟹的足部肌肉组织提取基因组DNA,构建Genome Walker Libraries,克隆C型凝集素(EsCTL)的基因组和启动子序列。结果显示,EsCTL的基因组全长为2 932 bp,其中结构基因由5个外显子(分别为33 bp33 bp165 bp57 bp157 bp)和4个内含子(分别为299 bp542 bp199 bp453 bp)构成;启动子区域包含4个启动子核心序列,6TBPTATA框结合蛋白),11C/EBPα(CCAAT增强子结合蛋白)和1SRF(血清应答因子)等真核生物的典型启动子结构特征,此外还包括15SP1(特异性蛋白1),2AP1(激活蛋白1),8GATA110Oct1等参与免疫调节的作用元件。EsCTL基因组及其启动子的克隆与特征分析,为深入研究EsCTL基因在免疫应答中表达调控奠定了基础。

关键词: 中华绒鳌蟹; 凝集素基因; 启动子; 克隆; 序列分析

中图分类号:  X 701文献标志码: A

引用格式:李凤梅, 祝倩倩. 中华绒螯蟹C型凝集素基因组和启动子的克隆与分析\[J\]. 青岛科技大学学报(自然科学版), 2018 393): 5359.

LI Fengmei ZHU Qianqian. Cloning and analysis of genome and promoter of Clectin gene from Eriocheir sinensis\[J\]. Journal of Qingdao University of Science and TechnologyNatural Science Edition), 2018, 39(3) 5359.

Cloning and Analysis of Genome and Promoter of

CLectin Gene from Eriocheir Sinensis

 

LI Fengmei ZHU Qianqian

(College of Marine Science and Biological Engineering Qingdao University of Science and Technology Qingdao 266042 China)

 

Abstract: In order to explore the transcriptional regulation mechanisms of lectin gene promoter from Chinese mitten crab Eriocheir sinensis in immune response, the Genome Walker Libraries from extracting genomic DNA of E. sinensis foot muscle tissue were constructed for cloning genome and promoter sequences of E. sinensis lectin (EsCTL) in this study. The results showed that the fulllength EsCTL genomic DNA was of 2 932 bp. The structural gene contained five exons (33 bp, 33 bp, 165 bp, 57 bp and 157 bp) and four introns (299 bp, 542 bp, 199 bp and 453 bp); the flanking sequence of 976 bp of EsCTL contained several typical transcription regulation elements, such as 4 promoter core sequences, 6 TBPs (TATAbox binding protein), 11 C/EBPαs (enhancer binding protein) and 1 SRF (serllm response factor), and several transcription regulation elements of immunerelated gene, such as 15 SP1sSpecificity Protein 1), 2 AP1s (activity protein 1), 8 GATA1s, and 10 Oct1s. The cloning and characterization of EsCTL genome and promoter provide references for the further study on expression regulation of EsCTL gene and its function analysis in the immune response.

Key words: Eriocheir sinensis; lectin gene; promoter; cloning; sequence analysis

 

收稿日期:    20170107

基金项目: 山东省高等学校科研计划项目(J13LE56.

作者简介: 李凤梅(1973—),女,副教授.

 

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