鸭瘟病毒基因组的提取和限制性酶切分析  

Extraction and Restriction Enzyme Mapping of Genomic DNA of Duck Plague Virus

作　　者：李玉峰[1] 何叶峰[2] 黄兵[2] 吴静[2] 于可响[2] 宋敏训[2] 杨汉春[1] 

机构地区：[1]中国农业大学农业部预防兽医学重点开放实验室,北京100094 [2]山东省农科院家禽研究所禽病研究中心,山东济南250023

出　　处：《中国家禽》2007年第5期12-14,共3页China Poultry

摘　　要：利用高渗缓冲液裂解感染鸭瘟病毒的鸭胚成纤维细胞,加入核酸酶消化细胞DNA,再抽提病毒DNA的方法提取鸭瘟病毒基因组,结果证明,这种方法可以最大限度地消除细胞DNA的污染,得到纯净的病毒基因组DNA。用10种限制性内切酶对鸭瘟病毒疫苗株和分离株的基因组进行酶切分析,酶切产物电泳结果表明,SmaⅠ、SalⅠ、PstⅠ、NotⅠ、HindⅢ、EcoRⅤ和BamHⅠ7种限制酶产生酶切图谱二者几乎完全一致,EcoRⅠ和SacⅠ仅有个别条带不同,BglⅡ产生的条带迁移率差别较大,但条带数量相同。Duck plague virus genomic DNA was prepared by combination of infected cells permeabilization by a Triton X-100 lysis buffer,digestion of cellular DNA by micrococcal nuclease,and subsequent extraction of viral DNA which is protected from nuclease by virion capsids. Nuclease treatment resulted in significant reduction of cellular DNA. The genomic DNA of vaccine strain and isolate SD-01 were digested by 10 restriction enzymes. The restriction enzyme maps of the two strain produced by SmaⅠ、SalⅠ、PstⅠ、NotⅠ、HindⅢ、EcoRⅤand BamHⅠare nearly identical. Maps produced by EcoRⅠand SacⅠdiffers by only few fragments. The mobility in the two maps produced by BglⅡ is much different,but the amount of the fragments is identical.

关 键 词：鸭瘟病毒 基因组DNA 提取 酶切图谱 

分 类 号：S83[农业科学—畜牧学]