H3N2犬流感病毒M1蛋白的原核表达及鉴定

Prokaryotic expression and preparation of H3N2 canine influenza virus M1 protein

关键词:  H3N2犬流感病毒  M1蛋白  原核表达  蛋白纯化

Key words:  H3N2 canine influenza virus  M1 protein  prokaryotic expression  protein purification

摘要:

对犬流感病毒(H3N2)M1基因进行克隆及原核表达,并制备出M1蛋白纯品。针对犬流感病毒(H3N2)M1基因序列设计引物,用聚合酶链式反应(PCR)扩增目的基因片段,扩增产物克隆至表达载体pET-SUMO中并转化至宿主菌BL21(DE3)。表达产物经一系列层析纯化及后续处理后,获得目的蛋白,并用Western blot检测纯化的M1目的蛋白。这项研究成功构建了pET-SUMO-M1并表达了M1蛋白,制备出的M1蛋白纯品为进一步制备通用型抗犬流感病毒抗体提供纯品抗原。

Abstract:

To prepare the high purity recombinant M1 protein, the canine influenza virus (H3N2) M1 gene was cloned and prokaryotically expressed. Primers targeting the canine influenza virus (H3N2) M1 gene were designed, then the H3N2 M1 gene was amplified by PCR and cloned into the expression vector pET-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3). The expressed product was purified by a series procedure, including chromatography and some subsequent treatment, then identified by Western blot. This study successfully constructed pET-SUMO-M1 and expressed the high purity recombinant M1 protein. The acquired high purity M1 protein provides a pure antigen for further preparation of a universal anti-canine influenza virus antibody.