Abstract: In this study, plasmid pEGFPN3 was introduced into the duck PGCs via lipofectamine in vitro. The transfected PGCs were injected into the subgerminal cavity of the recipient chicken to find a kind of method which would produce interspecies duck/chicken chimeras and contribute to transgenic study via chimeras. The Beijing duck was used as the donor and the Beijing Fatty chicken was the recipient, the duck/chicken chimeras were constructed via Windowing technique. The exogenous gene was expressed in PGCs transfected for 6 h later, and cultured for 24 h in vitro, the transfection rate reached 33.6%. 33 chicken embryos of 120 chicken eggs were obtained. Eventually 13 chimeric embryos were obtained, but there was no a surviving chicken. The exogenous gene were detected in 10.8% (13/120) of the embryos examined. Heterosexual cells were found in the gonads of 8 chimeras through PCR to amplify the Wspecific repeating sequences. The results indicated that it was possible to produce transgenic avians by constructing chimeras with the method adopted in this study. Furthermore, these results suggested that duck PGCs could migrate, colonize and proliferate in chicken gonads together with chicken′s PGCs during development.