Construction of recombinant pPIC9K-IL-2 and expression in Pichia pastoris

A pair of primers was designed to clone IL-2 gene according to the Ovis aries interleukin-2(IL-2)sequence and the recombinant expressed vector pPIC9K-IL-2 was constructed.The recombinane plasmid was transformed into Pichia pastoris GS115by electroporation.The transformants were selected with MD culture plate and identified by PCR.The multicopy recombinant P.pastoris strain was selected by G418(4g/L)resistance.The result showed that IL-2 gene was integrated with chromosome of P.pastoris by PCR identification.The expressed product had a molecular weight of 18ku band by SDS-PAGE analysis.The recombinant P.pastoris GS115 was successfully constructed,which provided the basis for research and development of a novel cytokine adjuvant.