| 合浦珠母贝表皮生长因子样(EGF-like)基因的克隆与表达分析 |
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| 引用本文： | 朱文杰,黄桂菊,张东玲,范嗣刚,刘宝锁,毕晓敏,喻达辉.合浦珠母贝表皮生长因子样(EGF-like)基因的克隆与表达分析[J].水产学报,2015,39(5):648-657. |
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| 作者姓名： | 朱文杰 黄桂菊 张东玲 范嗣刚 刘宝锁 毕晓敏 喻达辉 |
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| 作者单位： | 中国水产科学研究院南海水产种质资源与健康养殖重点实验室, 广东 广州 510300;中国水产科学研究院农业部南海渔业资源开发与利用重点实验室, 广东 广州 510300;中国水产科学研究院南海水产研究所, 广东 广州 510300;上海海洋大学水产与生命学院, 上海 201306,中国水产科学研究院南海水产种质资源与健康养殖重点实验室, 广东 广州 510300;中国水产科学研究院农业部南海渔业资源开发与利用重点实验室, 广东 广州 510300;中国水产科学研究院南海水产研究所, 广东 广州 510300,集美大学水产学院, 福建 厦门 361021,中国水产科学研究院南海水产种质资源与健康养殖重点实验室, 广东 广州 510300;中国水产科学研究院农业部南海渔业资源开发与利用重点实验室, 广东 广州 510300;中国水产科学研究院南海水产研究所, 广东 广州 510300,中国水产科学研究院南海水产种质资源与健康养殖重点实验室, 广东 广州 510300;中国水产科学研究院农业部南海渔业资源开发与利用重点实验室, 广东 广州 510300;中国水产科学研究院南海水产研究所, 广东 广州 510300,中国水产科学研究院南海水产种质资源与健康养殖重点实验室, 广东 广州 510300;中国水产科学研究院农业部南海渔业资源开发与利用重点实验室, 广东 广州 510300;中国水产科学研究院南海水产研究所, 广东 广州 510300;上海海洋大学水产与生命学院, 上海 201306,中国水产科学研究院南海水产种质资源与健康养殖重点实验室, 广东 广州 510300;中国水产科学研究院农业部南海渔业资源开发与利用重点实验室, 广东 广州 510300;中国水产科学研究院南海水产研究所, 广东 广州 510300 |
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| 基金项目： | 国家自然科学基金(31372525);现代农业产业技术体系建设专项(CARS-48);广东省海洋渔业科技推广专项(A201201A02,A201201A03,A201301A02,A201301A08,B201300B08);广东省科技计划(2012B050700004);中央级公益性科研院所基本科研业务费专项(2013YD04,2014TS08);海南省自然科学基金(312087) |
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| 摘 要： | 为探究表皮生长因子(epidermal growth factor,EGF)在合浦珠母贝幼体发育以及组织生长过程中的作用,实验通过RACE技术克隆了合浦珠母贝EGF-like基因并做了相应的表达分析。实验结果获得cDNA全长序列4 107 bp,命名为Pf-egf1,开放阅读框(ORF)702 bp,编码234个氨基酸,包含一个信号肽序列和一个跨膜结构域,功能结构域分析发现Pf-egf1具有一个EGF-like结构域,有6个半胱氨酸残基,由3个二硫键维持,是表皮生长因子家族及其相关蛋白的特征结构域,但其余序列与现有相关基因序列差异较大,推测可能是一个新的EGF-like基因。表达结果显示,Pf-egf1 mRNA在合浦珠母贝外套膜、闭壳肌、鳃、肝胰腺、珍珠囊、肠和性腺中均有表达,在肠中的表达显著高于其他组织;在合浦珠母贝幼体发育的担轮期、D型期、壳顶期、眼点期和变态期的表达呈现逐渐升高的趋势,并且其在变态期的表达量极显著地高于其他时期。上述结果表明,Pf-egf1基因可能在合浦珠母贝肠道修复和幼虫变态发育阶段起着重要作用,为进一步开展育珠与生长调控奠定了基础。
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| 关 键 词： | 合浦珠母贝 表皮生长因子 基因克隆 表达谱 |
| 收稿时间： | 2014/12/26 0:00:00 |
| 修稿时间： | 2015/2/25 0:00:00 |
Cloning and expression of an novel EGF-like gene from Pinctada fucata |
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| ZHU Wenjie,HUANG Guiju,ZHANG Dongling,FAN Sigang,LIU Baosuo,BI Xiaomin and YU Dahui.Cloning and expression of an novel EGF-like gene from Pinctada fucata[J].Journal of Fisheries of China,2015,39(5):648-657. |
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| Authors: | ZHU Wenjie HUANG Guiju ZHANG Dongling FAN Sigang LIU Baosuo BI Xiaomin and YU Dahui |
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| Institution: | Key Laboratory of South China Sea Fishery Genetic Resources and Health Aquaculture, Chinese Academy of Fishery Science, Guangzhou 510300, China;Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou 510300, China;South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China;College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China,Key Laboratory of South China Sea Fishery Genetic Resources and Health Aquaculture, Chinese Academy of Fishery Science, Guangzhou 510300, China;Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou 510300, China;South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China,Fisheries College, Jimei University, Xiamen 361021, China,Key Laboratory of South China Sea Fishery Genetic Resources and Health Aquaculture, Chinese Academy of Fishery Science, Guangzhou 510300, China;Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou 510300, China;South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China,Key Laboratory of South China Sea Fishery Genetic Resources and Health Aquaculture, Chinese Academy of Fishery Science, Guangzhou 510300, China;Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou 510300, China;South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China,Key Laboratory of South China Sea Fishery Genetic Resources and Health Aquaculture, Chinese Academy of Fishery Science, Guangzhou 510300, China;Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou 510300, China;South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China;College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China and Key Laboratory of South China Sea Fishery Genetic Resources and Health Aquaculture, Chinese Academy of Fishery Science, Guangzhou 510300, China;Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou 510300, China;South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China |
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| Abstract: | Epidermal growth factor(EGF)plays important roles in growth, wound healing and metamorphosis, etc. The pearl oyster Pinctada martensii is a commercially important species cultured for pearl production, in which a series of developmental transformation occur in young stage and an injury will happen during nuclear insertion. In order to evaluate the function of EGF gene during P. martensii larval development, tissue growth and nuclear insertion. In this study, cloning and expression of an EGF-like gene was carried out to provide some background for further study on growth development and pearl culture improvement. As a result, a full length cDNA of 4 107 bp was obtained and named Pf-egf1. The complete ORF encoded 234 amino acid residues, including a signal peptide, a transmembrane domain and an EGF-like domain. The EGF-like domain contained six cysteines that can form three disulfide bonds, consistent with the characteristics of Epidermal Growth Factor protein family. However, the remaining Pf-egf1 sequences were quite different from other EGF family members. So we speculated that Pf-egf1 is probably a novel EGF-like gene. The tissue expression profile showed that Pf-egf1 expressed in all the six tissues tested(mantle, muscle, gill, hepatopancreas, pearl sac, intestines and gonad), with significantly higher expression in intestines than in other tissues. The expression of Pf-egf1 presented an increasing trend from trochophore stage to metamorphosis stage in larval development, with significantly higher expression in metamorphosis stage than in other stages. The observations above indicated that Pf-egf1 gene may play important roles in intestine repairing and larval metamorphosis in P. martensii, possibly through promotion of cell proliferation, and thus can benefit to some extent further studies on modulation of growth and pearl culture. |
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| Keywords: | Pinctada martensii epidermal growth factor gene cloning expression profile |
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