Construction and expression of a site-specific mutant of human interleukin-2 gene in E.coli

Objective To construct expression clones of a site-specific mutagenesis(88N→R) of human interleukin-2(IL-2) gene to further investigate new kind of hIL-2 gene agent in clinical trial. Methods The human interleukin-2 cDNA containing full-length of encoding region was cloned by RT-PCR and confirmed by DNA sequencing. With recombinant PCR, authors amplified the whole genes of IL-2 with the site-directed mutagenesis and confirmed by DNA sequencing again, then cloned to the procaryotic expressive vector pGEX-4T-2 and transformated to E.coli.DH5 IL-2 recombinant clone was induced by IPTG to express the allosteric IL-2 proteins and the products were identified by SDS-PAGEPAGE.Results The mutated IL-2 gene clone was obtained successfully, while the recom-binant IL-2 proteins were efficiently expressed in E.coli DH5 Human IL-2 cDNA with the site-directed mutagenesis were expressed in E.coli, which has provided useful information for development of low-toxic, high-efficiency IL-2 recombinant agent.