Prokaryotic expression,purification of recombinant mouse interleukin-28 and production of its polyclonal antibody

Objective: To obtain purified recombinant interleukin-28 of mouse(mIL-28) and prepare its polyclonal antibody.Methods: The cDNA fragment coding for mature mIL-28 protein was amplified by PCR and cloned into vector pET-30 to construct fusion expression vector pET-30a-mIL-28.After pET-30a-mIL-28 was transformed into E.coli BL21(DE3),the bacteria were induced by IPTG.The expressed mIL-28 fusion protein was purified by Ni-NTA affinity chromatography.Six to eight weeks old chickens were immunized with the purified protein for obtaining the antiserum.The titers of antibodies were measured by ELISA. Results: The DNA sequencing showed that the expression vector pET-30a-mIL-28 was constructed successfully.After induced by IPTG,the expressed mIL-28 fusion protein in E.coli cultured at 37℃ appeared a single band on SDS-PAGE.The result of ELISA indicated that the prepared polyclonal antibody had high titer and specificity. Conclusion: The purified recombinant interleukin-28 of mouse and polyclonal antibody have been acquired.