| 三角帆蚌肝脏瘟病感染期抑制性消减cDNA文库的构建与分析 |
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| 引用本文： | 肖调义,葛熹凯,许宝红,苏建明,章怀云. 三角帆蚌肝脏瘟病感染期抑制性消减cDNA文库的构建与分析[J]. 水产学报, 2009, 33(5): 856-864 |
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| 作者姓名： | 肖调义 葛熹凯 许宝红 苏建明 章怀云 |
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| 作者单位： | 1. 湖南农业大学动物科技学院,湖南,长沙,410128
2. 湖南农业大学动物科技学院,湖南,长沙,410128;中南林业科技大学生命科学与技术学院,湖南,长沙,410004 |
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| 基金项目： | 湖南省科技厅攻关项目（06NK3049）；湖南省教育厅重点项目（07A012） |
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| 摘 要： | 采用抑制性消减杂交技术构建了三角帆蚌肝脏瘟病感染期抑制性消减cDNA文库。经检验，差异表达基因均被富集了 210倍左右，证明构建的 cDNA 消减文库具有很强的消减效率。PCR 鉴定发现，在随机挑取的阳性克隆中，95% 的克隆均含有 0.2～1.0 kb 的插入片段，这些片段可能是三角帆蚌瘟病病毒感染后差异表达基因的cDNA片段。测序共获得 214 个有效 cDNA 序列，分别属于8 大类，共 98 个基因。其中细胞分裂基因 2个、细胞结构与运动基因 9 个、代谢基因 10 个、信号传导基因 7 个、细胞防疫基因 10 个、基因与蛋白表达基因 20 个、未知功能蛋白基因 26 个，GenBank中找不到任何同源序列的基因 14 个。结果说明，构建的差异表达cDNA文库，可较好地反映三角帆蚌瘟病病毒对三角帆蚌影响的基因信息。
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| 关 键 词： | 三角帆蚌；肝脏；三角帆蚌瘟病病毒；抑制性消减杂交；消减cDNA文库 |
| 收稿时间： | 2008-09-02 |
| 修稿时间： | 2008-12-17 |
Construction and analysis of subtracted cDNA library by suppression subtractive hybridization from Hyriopsis cumingii liver |
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| XIAO Tiaoyi,GE Xikai,XU Baohong,SU Jianming and ZHANG Huaiyun. Construction and analysis of subtracted cDNA library by suppression subtractive hybridization from Hyriopsis cumingii liver[J]. Journal of Fisheries of China, 2009, 33(5): 856-864 |
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| Authors: | XIAO Tiaoyi GE Xikai XU Baohong SU Jianming ZHANG Huaiyun |
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| Affiliation: | Hunan Agricultural University |
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| Abstract: | This study used suppression subtractive hybridization (SSH) for construction of the cDNA subtractive library of the liver from the living body infected Hyriopsis cumingii plague virus. After testing, differentially expressed genes were enriched in the 210 times that the proof of the cDNA library has strong subtractive efficiency. Three hundred positive clones were randomly picked and identified by PCR method; 95% clones contained 0.2-1.0 kb by inserts,which might be the cDNA frag menu of differentially expressed genes in Hyriopsis cumingii plague virus infected group. Three hundred clones were sequenced and obtained 214 known function EST for 58 different genes were reported in H.cumingii for the first time in this study. According to the classification of functional genes by Adam,the identified EST in H.cumingii fell into eight categories relevant to 2 belong to cell division genes, 9 belong to cellular structure and movement genes, 10 belong to metabolism genes, 7 belong to signal transduction genes,10 belong to cell immune action genes, 20 belong to gene and protein expression associates genes and 26 belong to other proteins with unknown functions.In addition,there are 14 no similar sequence in Genbank, conjecturing its new genes. Results show, the method to construct cDNA subtractive library could well reflect the gene information of abalone affected by H. cumingii plague virus. Results of the present study provide the basic data to research of relationship between resistance breeding and gene expression of liver. |
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| Keywords: | Hyriopsis cumingii liver Hyriopsis cumingii plague virus suppression subtractive hybridization subtracted cDNA library |
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