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[1]陈玉磊,李婉玉,吴裕玲,等.皱纹盘鲍基质金属蛋白酶基因的克隆与表达分析[J].集美大学学报(自然科学版),2020,25(1):16-25.
　CHEN Yulei,LI Wanyu,WU Yuling,et al.Molecular Cloning and Expression Analysis of Matrix Metalloproteinases from Abalone(Haliotis discus hannai)[J].Journal of Jimei University,2020,25(1):16-25.
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《集美大学学报（自然科学版）》[ISSN:1007-7405/CN:35-1186/N]

卷:
第25卷
期数:
2020年第1期
页码:
16-25
栏目:
水产、食品与生物工程
出版日期:
2020-01-28

文章信息/Info

Title:
Molecular Cloning and Expression Analysis of Matrix Metalloproteinases from Abalone(Haliotis discus hannai)
作者:
陈玉磊1; 李婉玉1; 吴裕玲1; 杨汝晴1; 章骞1; 2; 张凌晶1; 刘光明1; 2; 曹敏杰1; 2
(1.集美大学食品与生物工程学院,福建 厦门 361021;2.水产品深加工技术国家地方联合工程研究中心,福建 厦门 361021)
Author(s):
CHEN Yulei1; LI Wanyu1; WU Yuling1; YANG Ruqing1; ZHANG Qian1; 2; ZHANG Lingjing1; LIU Guangming1; 2; CAO Minjie1; 2
(1.College of Food and Biological Engineering,Jimei University,Xiamen 361021,China2.National & Local Joint Engineering Research Center of Processing Technology for Aquatic Products,Xiamen 361021,China)
关键词:
基质金属蛋白酶; 皱纹盘鲍; 副溶血弧菌; 基因克隆; 荧光定量PCR
Keywords:
matrix metalloproteinases; Haliotis discus hannai; Vibrio parahemolyticus; gene cloning; quantitative real-time PCR
分类号:
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DOI:
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文献标志码:
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摘要:
为了探索基质金属蛋白酶（matrix metalloproteinases，MMPs）在软体动物中的存在形式及其在机体抗菌免疫中的作用，以皱纹盘鲍（Haliotis discus hannai）为研究对象，采用分子生物学技术克隆得到皱纹盘鲍MMP-16、MMP-17和MMP-21 3个基因部分序列，并对其进行生物信息学分析和多序列比对。此外，利用荧光定量PCR技术检测了Hdh-MMPs在皱纹盘鲍不同组织及副溶血弧菌感染前后各组织中的表达情况。结果显示，克隆得到的Hdh-MMPs序列包含完整的催化结构域和（或）类血红素结合区，Hdh-MMPs蛋白与多种软体动物的MMPs具有序列相似性。荧光定量PCR结果显示，Hdh-MMP-16和Hdh-MMP-17在血淋巴细胞中表达量最高，而Hdh-MMP-21在鳃中表达量最高。副溶血弧菌感染后，Hdh-MMP-16的表达量在鳃中呈现上升趋势，而Hdh-MMP-17的相对表达量呈现先上升后下降的趋势，Hdh-MMP-21基因在鳃和肝胰腺中呈现一定程度的上调表达。本研究结果表明，Hdh-MMP-16/17/21可能与皱纹盘鲍抵御病原菌入侵的先天免疫性调控相关。
Abstract:
In order to explore the existence of matrix metalloproteinases (MMPs) in molluscs and their role in the antibacterial immunity of the organisms,partial sequences of three MMP genes from Haliotis discus hannai,named as Hdh-MMP-16/17/21 were cloned,and the bioinformatics analysis and multiple sequence alignment were performed.In addition,the relative expression levels of Hdh-MMPs in different tissues of Haliotis discus hannai were preliminarily tested using quantitative real-time PCR (qPCR) before and after Vibrio parahaemolyticus infection.The results showed that Hdh-MMP-16/17/21 contained the complete catalytic domain and (or) hemopexin domain,and they presented a varying extent of sequence similarities with MMPs of various molluscs.The results revealed that the transcript levels of Hdh-MMP-16 and Hdh-MMP-17 were highest in hemocytes,while that of Hdh-MMP-21 was highest in gill.After Vibrio parahaemolyticus infection,the expression of Hdh-MMP-16 increased significantly in gill during the trial,while the relative expression of Hdh-MMP-17 increased first and then decreased in gill and hepatopancreas.The Hdh-MMP-21 gene expression was upregulated in gill and hepatopancreas to some extent post infection.Hence,it was shown that Hdh-MMP-16/17/21 might be involved in the innate immune responses to pathogen infection in abalone.

参考文献/References:

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备注/Memo

备注/Memo:
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更新日期/Last Update: 2020-04-17