Prokaryotic Expression and Preparation of Polyclonal Antibody for Porcine Interleukin-2 and Interleukin-6

The interleukin-2(IL-2) and interleukin-6(IL-6) are the important factors for immunoloregulation,and have great development prospects in the application of adjuvants.The genes of porcine IL-2 and IL-6 were amplified from recombinant plasmid pTIL-2 and pTIL-6.After being digested by restriction endonucleases,the IL-2 and IL-6 genes were subcloned into the prokaryotic expression vector pET30a.After restriction enzyme digestion,PCR identification and sequence analysis of the recombinant plasmids,the positive recombinant plasmids were selected and designated as pETIL-2 and pETIL-6.The pETIL-2 and pETIL-6 were transformed into E.coli BL21(DE3) and Rosetta(DE3) and then induced with IPTG at 37 ℃.SDS-PAGE and Western-blotting analysis showed that the expressed recombinant IL-2 and IL-6 proteins presented in inclusion bodies and were about 26ku and 30ku in molecular weight.The recombinant IL-2 and IL-6 protein which were purified and refolded were used to immunize rabbit for specific IL-2/IL-6 antibody generation.