论文部分内容阅读
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 establish a method for efficient purification of CP-CpG-DNA and to lay the foundation for the study of DNA immunopotentiating adjuvant function. Methods: Genomic DNA (CP-CpG-DNA) was extracted from Corynebacterium parvum (CP) by sonication, digestion with proteinase K, precipitation with CTAB, phenol and chloroform extraction. Results: The concentration of CP-CpG-DNA was 2.3Mb. The CP-CpG-DNA was digested with restriction endonuclease HpaII and detected by agarose gel electrophoresis. The fragment less than 1000bp was obtained. CONCLUSION: The CP-CpG-DNA obtained by the method of highly purified CP-CpG-DNA has good purity and a large amount of non-methylated CpG sites in CP-CpG-DNA, which can be used in DNA immunoadjuvant adjuvant .