不同方法提取鳗鲡病原菌DNA模板的差异分析  

Differentiation Analysis of the Templates DNA Extracted by Different Methods from Three Pathogenic Bacteria Strains of Eels

作　　者：熊静[1,2,3] 关瑞章[1,2,3] 郭松林[1,2,3] 黄文树[1,2,3] 关淑芳[1] 

机构地区：[1]集美大学水产学院,福建厦门361021 [2]鳗鲡现代产业技术教育部工程研究中心,福建厦门361021 [3]农业部东海海水健康养殖重点实验室,福建厦门361021

出　　处：《集美大学学报（自然科学版）》2012年第3期161-166,共6页Journal of Jimei University：Natural Science

基　　金：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)

摘　　要：利用吸光值、琼脂糖电泳及PCR差异扩增等指标,对酚-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16S rD-NA和外膜蛋白的保守序列.其中,酚-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16S rDNA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.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 bacte- ria strains isolated from eels by three methods of water boiling, alkali lyase and phenol-chloroform. The re- sults indicated that the templates DNA presented some differences in extracted concentration, purity and mole- cules weights; and the DNA extracted by the phenol-chloroform method has lower concentration, higher puri- ty 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 bac- teria strains.

关 键 词：模板DNA 水煮法 碱裂解法 酚-氯仿提取法 16SrDNA 外膜蛋白基因 

分 类 号：S851.347.34[农业科学—预防兽医学]