猪α干扰素原核表达载体的构建及表达

Gene Clonging and Prokarotic Expression of Porcine Interferon-α

以重组克隆质粒pGEM-T-IFN-αZ为模板,PCR扩增杂种猪α干扰素,将其插入原核表达载体pET-43.1a-c(+)中,构建了猪α干扰素原核表达载体pET-IFN-α,实现了猪α干扰素在大肠杆菌BL21中的表达.SDS-PAGE电泳、Western-blotting检测均出现了特异性的条带.SDS-PAGE电泳分析表明,表达的融合蛋白以可溶性形式存在.经薄层扫描分析,表达产物约占菌体总蛋白的29.8%.

Using recombination cloning plasmid pGEM-T-IFN-αZ as templat, porcine Interferon-alpha gene of crossbred growing and finishing pig was cloned. Interferon-alpha gene was inserted into pET-43, 1a-c （ ＋ ） of prokaryotic expression vector, and we obfained recombinant expression vector named pET-IFN-α. The recombinant expression vector can express fusion proteins of recombinant PoIFN-α in BL21 of E. coll. Specific strip appears on gel in SDS-PAGE and on NC-Membran in Western blotting. The result of SDSPAGE manifests that the target protein was expressed in soluble form. Aanalysed with thin slice scan, the expression products occupy 29.8 % in the whole thallus proteins.