短小棒状杆菌CpG-DNA提取方法的研究  

Studied on the method of extracting CpG-DNA from Corynebacterium parvum

作　　者：孙彬裕[1,2] 高尚先[2] 

机构地区：[1]山东大学免疫学研究所,济南250012 [2]中国药品生物制品检定所,北京100050

出　　处：《药物分析杂志》2009年第3期448-451,共4页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立一种高效纯化CP-CpG-DNA的方法,为研究DNA免疫增强佐剂功能奠定基础。方法:应用经超声破碎、蛋白酶K消化、CTAB沉淀及酚、氯仿抽提等步骤从短棒杆菌(Corynebacterium parvum,CP)中提取基因组DNA(CP-CpG-DNA)。结果:琼脂糖凝胶电泳检测,CP-CpG-DNA主要集中在2.3Mb;CP-CpG-DNA经限制性内切酶HpaII酶切后,琼脂糖凝胶电泳检测,得到小于1000bp的片断。结论:建立的高效纯化CP-CpG-DNA方法,获得的CP-CpG-DNA纯度较好,而且CP-CpG-DNA中存在大量非甲基化CpG位点,可用于DNA免疫增强佐剂的应用。Objective：To develop a efficient method for purification of CP-CpG-DNA for the study of DNA immune-enhancing adjuvant.Method：High quality of Corynebacterium parvum（CP）genome DNA（designated as CP-CpG-DNA）were prepared from the CP strain suspension by a series of treatments including sonification,protease k digestion,CTAB precipitation,and premixed phenol：chloroform：isoamyl alcohol extraction.Results：The extracted CP-Genomic-DNA has a molecular weight ranging from 1000-23130bp with a main band at about 23130bp by agarose gel electrophoresis analysis.The CP-CpG-DNA can be digested by HpaII enzyme to little fragments of less than 1000bp in length by agarose gel electrophoresis analysis,demonstrating hypersensitivity to HpaII,indicating high quantities of unmethylated CpG motifs in CP-CpG-DNA.Conclusion：It is a results give us a efficient method for purification of CP-CpG-DNA of CP.

关 键 词：短小棒状杆菌 基因组提取 HpaⅡ CPG基序 

分 类 号：R917[医药卫生—药物分析学]