鸭瘟病毒基因组核酸的提取方法比较  

COMPARISON OF EXTRACTION METHOD OF GENOMIC DNA OF DUCK PLAGUE VIRUS

作　　者：李子剑[1] 李玉峰[2] 陈苏[1] 宋敏训[2] 李云龙[1] 

机构地区：[1]山东师范大学生命科学学院动物抗性生物学重点实验室,济南250014 [2]山东省农业科学院家禽研究所禽病诊断与免疫重点实验室,济南250023

出　　处：《山东师范大学学报（自然科学版）》2008年第1期134-136,共3页Journal of Shandong Normal University(Natural Science)

基　　金：山东省农业科学院青年科研基金项目(2005YQ0037)资助

摘　　要：将鸭瘟病毒分离株SD-01在鸡胚成纤维细胞上增殖.收集病变细胞及培养液,采用两种方法提取病毒基因组核酸.第一种方法,首先用差速离心的方法纯化病毒粒子,然后用酚/氯仿抽提的方法提取病毒基因组.第二种方法,先用高渗缓冲液将感染DPV的细胞完全裂解,然后加入微球菌核酸酶消化细胞DNA,最后用酚/氯仿抽提.限制性内切酶EcoRⅠ消化,结果表明,第一种方法提取的样品中含有细胞DNA,酶切后为弥散模糊的拖带,而方法二可以最大限度的消除细胞DNA污染,得到纯净的病毒基因组DNA,酶切后为清晰的梯状条带.The SD-01 strain of DPV was propagated in chicken embryo fibroblast monolayer cells.The genomic DNA of DPV was extracted by two methods.The first was to purify virons first and then obtain genomic DNA by phenol/chloroform extraction.The second was combination of infected cells permeabilization by a Triton X-100 lysis buffer,digestion of cellular DNA by Micrococcal nuclease and subsequently extraction of the viral DNA with the phenol/chloroform.The digestion of restriction enzyme EcoR I indicated that the nuclease treatment resulted in significant reduction of cellular DNA.

关 键 词：鸭瘟病毒 基因组核酸 微球菌核酸酶 

分 类 号：Q939.47[生物学—微生物学]