IL-18-PE38真核表达质粒的构建及表达

Construction and Expression of the Eukaryotic Expression Plasmid of Interleukin-18-PE38

目的构建含有白细胞介素18(Interleukin18,IL-18)及毒素融合基因的真核表达质粒,并研究其在NIH3T3细胞中的表达情况。方法通过分子克隆技术,将IL-18-PE38(Pseudomonas exotoxin A)融合基因插入真核表达载体Psec Tag2B中,构建真核表达质粒Psec Tag2B-IL-18-PE38,重组质粒经限制性内切酶及PCR测定证实构建成功后,用脂质体介导法将其转染小鼠NIH3T3细胞中,然后用免疫荧光技术鉴定其表达。结果经酶切及PCR鉴定证实IL-18-PE38融合基因已被正确地插入真核表达载体Psec Tag2B中。荧光免疫细胞化学法荧光显微镜照片证实此重组基因可在其中表达。结论成功构建了重组的真核表达载体Psec Tag2B-IL-18-PE38并在体外细胞株获得表达,为下一步动物实验打下基础。

Objective To construct the IL-18-PE38 fusion gene expression plasmid and explore its expression in NIH3T3 cells.Methods The interleukin-18-PE38 fusion gene was cloned into the eukaryotic expression vector PsecTag2B to get the recombinant plasmid PsecTag2B-IL-18-PE38 by molecular cloning technique.It was transferted into NIH3T3-cell lines with liposome after it had been confirmed by restrictive enzyme analysis and PCR.Results Restrictive enzyme analysis and PCR demonstrated that the interleukin-18-PE38 fusio...