Cloning and expression of gushi-chicken interleukin 2 gene and detection of its biological activity

Two pairs of primers of RT-PCR and expressing were designed and synthesied according to the published gene sequence of chicken interleukin-2. About 680 bp target DNA sequence were cloned by RT-PCR depending on the template of total RNA isolated from ConA-stimulated spleen cell and inserted into the plasmid pGEM-T. The sequence coded ChIL-2 was subcloned to the pET28a vector. The expressing plasmid, recombined pET28a-ChIL-2, identified by enzyme digecting and DNA sequencing , was transformed into BL21(DE3) and induced with IPTG. SDS-PAGE illuminated that the expressed protein was 18 ku. recombinant protein was extracted roughly from dissolved BL21,and purified by Ni+ affinity column. MTT colorimetric assay indicated that the recombinant protein could induce chicken spleen T lymphocytes in vitro. Prepression was made for the advanced reserch of the usage of recombinant IL-2 protein as adjuvant to accine and antigene to monoclonal antibody.