鸡传染性支气管炎病毒四川分离株N基因DNA免疫质粒的构建  

DNA vaccine plasmid construction of N gene of avian infectious bronchitis SAIB_(WJ) strain isolated in Sichuan

作　　者：马孟根[1] 王红宁[2] 余勇[3] 李春[3] 周生[2] 杨帆[2] 刘世贵[1] 

机构地区：[1]四川大学生命科学学院,四川成都610041 [2]四川农业大学动物科技学院,四川雅安625014 [3]四川省动物防疫监督总站,四川成都610041

出　　处：《中国兽医科技》2005年第1期17-21,共5页Chinese Journal of Veterinary Science and Technology

基　　金：国家高技术研究发展计划(863)项目(2003AA241120)

摘　　要：采用SDS 蛋白酶K法抽提鸡传染性支气管炎病毒(IBV)四川分离株SAIBWJ的基因组RNA,参照基因库中的N基因序列设计了1对引物,扩增出了单一的DNA产物。将该产物插入pUC18载体的HindⅢ和BamHⅠ酶切位点,构建了pUC NWJ质粒。序列测定结果表明,获得的克隆质粒包含了全部N基因。再将N基因亚克隆到pcDNA3.1(+)载体的HindⅢ和XhoⅠ酶切位点,构建了IBVSAIBWJN基因的DNA免疫质粒pcDNA NWJ。The genome RNA of IBV strain SAIB_(WJ)isolated in Sichuan was extracted by SDS-proteinase K method. A pair of primers was designed and synthesized according to the sequences of N gene of IBV in GenBank. A single strip DNA amplified by PCR was inserted into pUC18 vector in HindⅢ and BamHⅠ sites, and the recombinant plasmid of pUC-N_(WJ)was constructed. The sequencing result showed that the cloning plasmid contained the full length of N gene. The N gene was sub-cloned into pcDNA3.1(+) plasmid in HindⅢ and XhoⅠ sites, and the pcDNA-N_(WJ)plasmid of N gene of IBV SAIB_(WJ)strain isolated in Sichuan was constructed.

关 键 词：DNA免疫 质粒 UC 基因组RNA 分离株 PCDNA3 扩增 N基因 鸡传染性支气管炎病毒 IBV 

分 类 号：S852[农业科学—基础兽医学] R373[农业科学—兽医学]