不同方法提取鳗鲡病原菌DNA模板的差异分析-《集美大学学报（自然科学版）》

文章信息/Info

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; (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)

Keywords:: template DNA; water-boiling method; alkaline lyase method; phenol-chloroform method; 16S rDNA; OMP gene

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 rDNA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.

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.