| 红笛鲷头肾消减cDNA文库的构建与分析 |
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| 引用本文： | 张新中,吴灶和,简纪常,鲁义善.红笛鲷头肾消减cDNA文库的构建与分析[J].中国水产科学,2011,18(2):379-391. |
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| 作者姓名： | 张新中 吴灶和 简纪常 鲁义善 |
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| 作者单位： | 1. 中国科学院南海海洋研究所,广东,广州,510301;中国科学院研究生院,北京,100049;广东省水产经济动物病原生物学及流行病学重点实验室,广东,湛江,524025;广东省教育厅水产经济动物病害控制重点实验室,广东,湛江,524025
2. 广东省水产经济动物病原生物学及流行病学重点实验室,广东,湛江,524025;广东省教育厅水产经济动物病害控制重点实验室,广东,湛江,524025 |
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| 基金项目： | 国家自然科学基金资助项目，广东省科技厅国际合作项目 |
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| 摘 要： | 应用抑制性消减杂交技术(SSH)构建红笛鲷(Lutjanus sanguineus)头肾消减cDNA文库,筛选红笛鲷免疫相关基因的EST.以哈氏弧菌(Vibrio harveyi)灭活疫苗体内诱导红笛鲷为实验组,以注射无菌生理盐水的红笛鲷为驱动组,通过SSH技术构建红笛鲷头肾消减.DNA文库.利用PCR技术和斑点杂交对文库进行筛选,从2 424个含插人片段的阳性克隆中筛选了680个克隆在上海生工进行了序列测定.使用BLASTx和BLASTn工具对获得的ESTs与GenBank数据库进行同源性比较并根据相似性序列的名称通过GO法对ESTs进行注释.结果获得了30个与红笛鲷免免疫防御相关基因的EST,如组织相容性抗原复合物基因(MHC I和MHCII),免疫球蛋白基因(IgH和IgL)、热休克蛋白基因(HSP10,HSP70和HSP90)等.本研究构建了哈氏弧菌灭活疫苗免疫后与正常组织差异表达的消减cDNA文库,并获得一批与红笛鲷免疫防御相关的ESTs,旨在为探讨红笛鲷分子免疫防御机制、筛选参与免疫防御调控相关的功能基因,揭示红笛鲷免疫抗病机制、提高机体抗病力、实现遗传改良奠定基础.
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| 关 键 词： | 红笛鲷 抑制性消减杂交技术 cDNA文库 免疫基因 |
| 修稿时间： | 2011/3/9 0:00:00 |
Construction and analysis of subtractive cDNA library of head kidney in humphead snapper,Lutjanus sanguineus |
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| ZHANG Xinzhong,WU Zaohe,RAN Jichang,LU Yishan.Construction and analysis of subtractive cDNA library of head kidney in humphead snapper,Lutjanus sanguineus[J].Journal of Fishery Sciences of China,2011,18(2):379-391. |
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| Authors: | ZHANG Xinzhong WU Zaohe RAN Jichang LU Yishan |
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| Institution: | ZHANG Xinzhong 1,2,3,4,WU Zaohe 3,JIAN Jichang 3,LU Yishan 3,4 1.South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301,China,2.Graduate School of Chinese Academy of Sciences,Beijing100049,3.Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Zhanjiang 524025,4.Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals |
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| Abstract: | A subtracted cDNA library of humphead snapper (Lutjanus sanguineus) was constructed by suppression subtractive hybridization technology (SSH) to screen immune-related EST.The cDNA library has been constructed by the mRNA of the test group and driven by SSH.Differential ESTs from the subtracted cDNA library have been identified by both PCR technology and dot blot hybridization.Six hundred and eighty positive clones were sequenced by Sangon Biological Engineering Technology & Services Co.,Ltd.The homology of ... |
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| Keywords: | Lutjanus sanguineus suppression subtractive hybridization cDNA library immunologic gene |
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