Abstract: [Objective]To construct a eukaryotic expression vector to express chicken beta-defensin-2 （gal-2）.[Methods]With total RNA extracted from chicken lung tissues as a template,full-length gal-2 cDNA was obtained by RT-PCR.The gene of mature peptide obtained by PCR was cloned into the eukaryotic expression vector pPIC3.5K to yield recombinant plasmid pPIC3.5K-gal-2 which was identified by DNA sequencing.The pPIC3.5K-gal-2 was then transformed into the competent Pichia pastoris GS115 and induced by methanol.The recombinant gal-2 protein was analyzed by SDS-PAGE.[Results]The lined recombinant plasmid DNA was successfully transformed into the competent Pichia pastoris GS115 and integrated into GS115 chromosome through electroporation and it expressed target protein with methanol induction.[Conclusion]The eukaryotic expression vector pPIC3.5K-gal-2 was constructed,and the 4.1 kDa chicken gal-2 protein was expressed in Pichia pastoris.The research provided a basis for further large-scale production of gal-2 protein by genetic engineering and protein engineering techniques.