| 黄鳝脑芳香化酶基因cDNA的克隆及组织表达特异性分析 |
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| 引用本文： | 李伟,刘功银,罗春桃.黄鳝脑芳香化酶基因cDNA的克隆及组织表达特异性分析[J].水产科学,2009,28(8). |
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| 作者姓名： | 李伟 刘功银 罗春桃 |
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| 作者单位： | 长江大学,生命科学学院,湖北,荆州,434025 |
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| 基金项目： | 长江大学博士创新基金资助项目 |
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| 摘 要： | 根据NCBI数据库报道的黄鳝芳香化酶基因的gDNA序列,设计了一对特异性引物.用Trizol试剂盒提取黄鳝脑总RNA,反转录获得cDNA第一条链,进行PCR扩增.扩增产物经过回收、连接、测序后得到了一条长1514 bp的芳香化酶基因cDNA序列.同源性比较表明该基因属于脑型P450aromB,与其他鱼类脑型P450aromB的同源性较高(>70%),与性腺型P450aromA的同源性较低(<60%),与自身性腺型芳香化酶同源性为59.5%.采用RT-PCR的方法对该基因在雄性黄鳝的各组织表达情况进行了分析,结果表明,该基因的转录本较高的表达于脑和精巢中,较低的在皮肤中表达,而在肝脏、心脏、小肠、肌肉等组织中没有检测到该基因的表达.
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| 关 键 词： | 黄鳝 芳香化酶 克隆 表达 |
Molecular Cloning and Tissue-specific Expression Analysis of Brain P450arom in Ricefield Eel(Monopterus albus) |
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| LI Wei,LIU Gong-yin,LUO Chun-tao.Molecular Cloning and Tissue-specific Expression Analysis of Brain P450arom in Ricefield Eel(Monopterus albus)[J].Fisheries Science,2009,28(8). |
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| Authors: | LI Wei LIU Gong-yin LUO Chun-tao |
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| Institution: | College of Life Science;Yangtze University;Jingzhou 434025;China |
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| Abstract: | Based on one P450arom genomic DNA sequence(EU840259),a pair of gene-specific primers was designed and synthesized.After extraction of brain total mRNA using Trizol and cDNA synthesized with M-MLV,PCR was done to amplify the cDNA of the brain P450arom.Subsequently,the PCR product was purified and cloned into pMD-18T vector.Sequencing results showed that the rice field eel brain P450arom was 1514bp in length,containing the full open reading frame of the gene.Sequence and phylogenetic analysis revealed that th... |
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| Keywords: | Monopterus albus P450arom molecular cloning expression analysis |
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