Abstract: Objective To study the transfection efficiency of pSecTag in chicken embryonic fibroblast cells, obtained the optimum condition of transfection, and established the foundation for the target gene expression in embryonic fibroblast cells of chicken. Method EGFP gene was cloned by PCR and constructed secretive eukaryotic expression vector by digestion, ligation and transformation, the relative content of the green fluorescent protein were detected by experimental design optimization. Results The optimum condition of transfection was with 2μl lipofectamine and 1μg expression vector in medium. Conclusion This method can effectively transfect the chicken embryo fibroblast.

Key words: CEF, EGFP； Eukaryotic expression vector