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

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.