tPA信号肽和猪繁殖与呼吸综合征病毒ORF5基因原核表达载体的构建

根据GenBank（序列号E00896）上公布的人组织型纤溶酶原激活因子（tissue plasminogen activator，tPA）的序列，利用在线信号肽分析工具Signal P3.0 Server预测出人tPA的信号肽共有63 bp，设计1对能互为模板直接进行扩增的特异性引物，通过重叠PCR的方法扩增出tPA信号肽。另设计1对连接引物，利用PCR方法将tPA信号肽与已删除自身信号肽的ORF5基因相连，成功构建了用tPA信号肽取代猪繁殖与呼吸综合征病毒ORF5基因原有信号肽的原核表达质粒，为下一步GP5蛋白分泌表达及猪繁殖与呼吸综合征疫苗开发、诊断试剂盒的研制与应用奠定基础。

Construction of Prokaryotic Expression Vector of PRRSV ORF5 Gene and tPA Signal Peptide

According to the published gene sequence of tissue plasminogen activator(tPA)from GenBank(accession number E00896),with on-line signal peptide analysis tools to predict the tPA signal peptide including 63 bp,one pair of specific primer of tPA signal peptide was designed and synthesized and the tPA signal peptide was amplified by overlapping PCR.Finally,the tPA signal sequence was introduced to replace the original signal peptide of the ORF5 gene by another pair of primer.The studies lay foundations for furt...