摘要： 构建人源白介素（h-IL2）基因的表达载体,并尝试在原核细胞中诱导表达,以用于进一步重组蛋白纯化和抗体生产。以h-IL2基因为模板,采用PCR技术扩增h-IL2基因,连接到pMD-18T载体中构建克隆载体,转化到感受态DH5α,提取质粒PCR、双酶切验证连接成功,经DNA测序鉴定基因序列与GenBank数据库收录h-IL2基因（NM₀00586.3）序列一致;再将h-IL2基因插入到pGEX-4T-1构建表达载体,转化至感受态BL21（DE3）,筛选阳性克隆IPTG诱导。SDS-PAGE验证成功表达融合蛋白GST-IL2。

关键词: 白细胞介素, 基因克隆, 原核表达

Abstract: To construct a recombinant espression vector of human Interleukin-2,and eapress in prokaryotic cells,which is helpful for the further purification of recombinant proteins and antibodies production.Methods:Amplify the h-IL2 gene by PCR,the DNA fragments were cloned into pMD-18T simple vectors,transformed into competent DH5α,recombinant plasmid PCR,restriction enzyme digest and sequencing confirmed that the fragment was h-IL2 and the sequence was identical to that published in GenBank.(NM_000586.3).Then the h-IL2 gene were cloned into pGEX-4T-1 constructing expression vectors,transformed into competent BL21(DE3),induced by IPTG,SDS-PAGE verification successful expression of the fusion protein GST-IL2.

Key words: Human Interleukin, Gene cloning, Prokaryotic expression

中图分类号: 

• Q78