| 不同方法提取鳗鲡病原茵DNA模板的差异分析 |
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| 引用本文： | 熊静,关瑞章,郭松林,黄文树,关淑芳. 不同方法提取鳗鲡病原茵DNA模板的差异分析[J]. 集美大学学报(自然科学版), 2012, 17(3): 161-166 |
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| 作者姓名： | 熊静 关瑞章 郭松林 黄文树 关淑芳 |
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| 作者单位： | 1. 集美大学水产学院,福建厦门361021 鳗鲡现代产业技术教育部工程研究中心,福建厦门361021 农业部东海海水健康养殖重点实验室,福建厦门361021
2. 集美大学水产学院,福建厦门,361021 |
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| 基金项目： | 国家自然科学基金项目(31172438,31001136);福建省科技厅重点项目(2010N0022,2009N0046);福建省教育厅重点项目(JA09157);厦门市科技局重点项目(3502Z20103023) |
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| 摘 要： | 利用吸光值、琼脂糖电泳及PCR差异扩增等指标，对酚一氯仿法、水煮法、碱裂解法3种方法制备的鳗鲡病原菌DNA模板进行了差异分析．结果表明：1）不同方法提取的病原菌模板DNA的浓度、纯度及分子质量大小存在差异，但以这3种方法提取的病原菌DNA为模板，均能成功扩增到细菌16SrD．NA和外膜蛋白的保守序列．其中，酚一氯仿提取的模板DNA浓度较低、纯度较高；水煮法和碱裂解法提取的模板DNA浓度较高、纯度较低．2）电泳显示，3种方法获得的模板DNA扩增出的16SrDNA保守序列条带均一，没有显著差异，但在外膜蛋白保守序列的扩增中却因菌种和模板提取方法的不同而存在差异．
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| 关 键 词： | 模板DNA 水煮法 碱裂解法 酚-氯仿提取法 16S rDNA 外膜蛋白基因 |
Differentiation Analysis of the Templates DNA Extracted by Different Methods from Three Pathogenic Bacteria Strains of Eels |
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| XIONG Jing,GUAN Rui-zhang,GUO Song-lin,HUANG Wen-shu,GUAN Shu-fang. Differentiation Analysis of the Templates DNA Extracted by Different Methods from Three Pathogenic Bacteria Strains of Eels[J]. the Editorial Board of Jimei University(Natural Science), 2012, 17(3): 161-166 |
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| Authors: | XIONG Jing GUAN Rui-zhang GUO Song-lin HUANG Wen-shu GUAN Shu-fang |
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| Affiliation: | 1(1.Fisheries College,Jimei University,Xiamen 361021,China;2.Engineering Research Center of Eel Industrial Technology,Ministry of Education,Xiamen 361021,China;3.Key Laboratory of Healthy Mariculture for East China Sea,Ministry of Agriculture,Xiamen 361021,China) |
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| Abstract: | By the UV absorbance values,maps of agarose gel electrophoresis and the PCR amplification results,this study analyzed the differentiations of those templates DNA extracted from three pathogenic bacteria strains isolated from eels by three methods of water boiling,alkali lyase and phenol-chloroform.The results indicated that the templates DNA presented some differences in extracted concentration,purity and molecules weights;and the DNA extracted by the phenol-chloroform method has lower concentration,higher purity and smaller molecules weights comparing with those DNA templates extracted by the other two methods.And all the templates DNA showed no difference in amplifying the conserved sequences of 16S rDNA,while the showed some differences in amplifying the partial out membrane protein(OMP)gene from the three bacteria strains. |
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| Keywords: | template DNA water-boiling method alkaline lyase method phenol-chloroform method 16S rDNA OMP gene |
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