| TAIL-PCR方法克隆约氏黄杆菌aroA基因序列（摘要） |
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| 引用本文： | 刘礼辉,李宁求,石存斌,吴淑勤.TAIL-PCR方法克隆约氏黄杆菌aroA基因序列（摘要）[J].农业科学与技术,2010,11(7):179-182. |
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| 作者姓名： | 刘礼辉 李宁求 石存斌 吴淑勤 |
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| 作者单位： | 中国水产科学研究院珠江水产研究所,广东广州,510380 |
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| 基金项目： | The Natural Science Foundation of Guangdong Province，National Special Fund for the Agricultural Commonweal Industry，Earmarked Fund for Modern Agroindustry Technology Research System(nycytx-49-14).广东省自然科学基金项目(06024033).农业公益性行业科研专项，现代农业产业技术体系建设专项基金 |
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| 摘 要： | 目的]研究草鱼烂鳃病病原约氏黄杆菌芳香氨基酸代谢途径的合成基因aroA基因的全序列。方法]以草鱼烂鳃病病原M165菌株基因组DNA为模板,分别利用5′和3′的3个特异性巢式引物与随机引物,通过热不对称交错PCR（TAIL-PCR）扩增菌株M165的aroA序列基因,并对其序列进行分析。结果]电泳检测结果表明,扩增产物与预期扩增结果相符;测序结果分析表明,aroA基因序列全长1230bp,编码410个氨基酸。aroA蛋白序列与Flavobacterium johnsoniae的aroA蛋白质序列,具有高度同源性。结论]TAIL-PCR方法为基因全序列的克隆提供了一种简便、高效的方法。aroA基因全序列的获得为进一步阐明aroA等营养相关因子对鱼类烂鳃病病原的致病力的影响奠定基础。
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| 关 键 词： | 热不对称PCR 烂鳃病 约氏黄杆菌 aroA基因 |
Efficient Amplification and Sequence Analysis of aroA Gene Sequence of Flavobacterium johnsoniae through TAIL-PCR |
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| LIU Li-hui,LI Ning-qiu,SHI Cun-bin,WU Shu-qin.Efficient Amplification and Sequence Analysis of aroA Gene Sequence of Flavobacterium johnsoniae through TAIL-PCR[J].Agricultural Science ＆ Technology,2010,11(7):179-182. |
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| Authors: | LIU Li-hui LI Ning-qiu SHI Cun-bin WU Shu-qin |
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| Institution: | ( Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380 ) |
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| Abstract: | Objective] The aim was to study the sequence of aroA gene （synthetic gene in metabolic pathway of aromatic amino acids） of a pathogenic bacterium （Flavobacterium johnsoniae） causing gill-rote disease.Method] Genomic DNA of strain M165 of F.johnsoniae was used as a template,three specific nested primers of 5＇end and 3＇end and arbitrary primer were used to amplify the aroA gene of strain M165 through Thermal asymmetric interlaced PCR （TAIL-PCR）.And the obtained sequence was analyzed.Result] The electrophoresis determination result showed that the size of amplified product was consist with the expected product; sequencing analysis suggested that the full-length of aroA gene was 1 230 bp,enconding 410 amino acids.The amino acid sequence of aroA protein showed the highest level of similarity to amino acids sequence of aroA protein of F.johnsoniae.Conclusion] TAIL-PCR provided a simple and efficient new method for the cloning of the gene sequence.Obtaining of full-length of aroA gene provided a foundation for further investigation on the effects of nutrition correlation factors such as aroA on the virulence of and virulence of F.johnsoniae. |
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| Keywords: | TAIL-PCR Gill-rote disease Flavobacterium johnsoniae aroA gene |
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