Chinese Journal of Veterinary Science 2005;25(2):178-182

Expression of Bovine Interleukin-2 Gene in Pichia pastoris

Fang CHEN ; Hongli SUN ; Xiangrong CAO ; Zhen LI ; Ruisong YU

Keywords

bovine Interleukin-2; Pichia pastoris; gene expression; pPICZB

Country

China

Language

Chinese

Abstract

The Interleukin-2 gene cDNA was cloned into the Pichia pastoris expression vector pPICZB,which is under the control of the alcohol oxidase promoter AOX1. The linearized recombinant plasmid of BoIL2-pPICZB,digested by Sac I ,was transformed into X-33 strains by electroporation. The multi-copy insert transformants were screened by Zeocin-resistance and induced by 1% methanol. The intracellular expression products were tested by SDS-PAGE analysis and Western blotting. Purified recombinant BoIL2 was gained by metal-chelating affinity chromatographic (MCAC). Assay with murine CTLL-2 cells showed that the recombinant BoIL2 exhibited the biological activity.