缺失信号肽和跨膜区的PRRSV GP5融合蛋白基因的表达  

Procaryotic expression of GP5 fusion protein gene deleted signal peptide and transmembrane region from PRRSV

作　　者：王明翠[1] 任晓峰[1] 李广兴[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2010年第4期342-346,共5页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30700590);黑龙江省自然科学基金重点项目(ZJN0702-01)

摘　　要：通过分析猪繁殖与呼吸综合征病毒(PRRSV)HH08株GP5基因序列,设计了2对引物,经PCR扩增,获得不含信号肽和跨膜功能区的GP5基因片段,长度分别为121 bp和248 bp。应用酶切位点相连构建融合基因技术,将2段基因亚克隆到原核表达载体pET-30a(+)中,获得重组表达质粒pET-GP5。将重组质粒转入大肠杆菌,经IPTG诱导后获得高效表达。免疫印迹试验证实,表达的融合蛋白能与特异性血清抗体反应,表明表达的PRRSV GP5蛋白具有良好的生物学活性。Using two pairs of primers,the N terminal and C terminal parts of GP5 gene were amplified from porcine reproductive and respiratory syndrome virus（PRRSV）.The signal peptide sequences and transmembrane regions in the N terminal and the C terminal parts of GP5 gene were deleted by such amplification.Two gene fragments,which were 121bp and 248bp in size respectively,were linked using three repeated peptide sequences and then cloned into the prokaryotic expression vector pET-30a（＋）.The resulting recombinant plasmid was transformed into Escherichia coli.The bacterially expressed GP5 protein was recognized by specific antibody,confirming that the protein had biological activity.

关 键 词：猪繁殖与呼吸综合征病毒 GP5基因 蛋白表达 

分 类 号：S852.659.6[农业科学—基础兽医学]