The cDNA encoding quail (coturnix) ovalbumin was cloned from quail oviduct by the method of RT-PCR, which was inserted into the P. pastoris genome downstream of the methanol inducible 5' alcohol oxidase (AOX) promoter to replace the AOX1 gene using the plasmid vector pPIC9, and a recombinant P. pastoris strain efficiently secreting quail ovalbumin was successfully constructed. ELISA analysis using a polyclonal antibody raised against quail ovalbumin showed that, induction by 0.75% methanol for 48h led to synthesis of secreted quail ovalbumin to give around 5.45 g l-1, respectively. The recombinant ovalbumin was purified into homogeneity later through ion exchange and gel filtration chromatography. SDS-PAGE analysis revealed that, compared to natural ovalbumin, the recombinant ovalbumin is glycosylated in the similar extent by P. pastoris.