鸭肠炎病毒us2基因转移载体的构建  

Construction of Transfer Vector of Duck Enteritis Virus us2 Gene

作　　者：刘晓玫[1] 赵妍[1] 乌伊罕[1] 张智慧[1] 马波[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国家禽》2009年第17期25-28,共4页China Poultry

基　　金：国家"十一五"科技支撑计划(2006BAD06A05-1)

摘　　要：以鸭肠炎病毒(DEV)C-KCE株基因组DNA为模板,PCR扩增得到us2全基因(包含s3、us3部分基因),将扩增产物克隆入pMD18-T载体,EcoRⅠ和HindⅢ双酶切连入pUC19载体,获得质粒pUC-us2。将完整的EGFP表达盒插入us2基因内BamHⅠ位点,得到转移载体质粒pUC-us2-EGFP。脂质体法将转移载体质粒pUC-us2-EGFP转染DEF细胞,观察到绿色荧光,说明EGFP基因获得表达。With the DNA of duck enteritis virus（DEV）-KCE as temple,us2 gene（including part of s3 and us3）was amplified by polymerase chain reaction （PCR）,then cloned into vector pMD18-T. The product was cloned into pUC19 vector digested by EcoR I and Hind III. Then the EGFP cassette was inserted into BamH I sites of the us2 gene to construct the plasmid of pUC-us2-EGFP. The transfer vector pUC-us2-EGFP was transfected into DEF cell with lipidosome,and the green fluorescence observed,the results showed that EGFP gene could be expressed.

关 键 词：鸭肠炎病毒 us2基因 转移载体 

分 类 号：S852.659.1[农业科学—基础兽医学] S858.31[农业科学—兽医学]