The Construction And Expression Of A Site-specific Gene Mutant Of The Human Interleukin-2 Gene In E.coli

Objects: To construct expressive clones of a site-specific mutagenesis(88N-R) of human interleukin-2(IL-2) gene with the aim of furthering the investigation of the new kind of hIL-2 genie drugs 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, We 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 a .The IL-2 recombinant clone was induced with 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, the recombinant IL-2 proteins were efficiently expressed in E.coli DH5 a . Conclusion: 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 drugs.