Objective:To construct an eukaryotic expression recombinant plasmid pEGFP-N1-PLZF and get its protein translation in spermatogonia of the neonatal rats in vitro. Methods:Using the primers of PLZF gene and the total RNA extracted from the tissue of rat testes, RT-PCR was performed. The product was inserted to multiple clone sites of EGFP-N1 vector by DNA recombinant technique. Then a new eukaryotic expression recombinant plasmid pEGFP-N1-PLZF was generated and identified by incision enzyme EcoRⅠ and SalⅠ and DNA sequencing. pEGFP-N1-PLZF was transfected into spermatogonia by liposome, from which protein were extracted after 24 h for detecting their expression by Western blot analysis. The treated cells were continuously traced by fluorescence microscope. Results:The recombinant plasmid cut by incision enzyme EcoRⅠ and SalⅠ overnight generated a 2 kb fragment,and DNA sequence of the 2 kb fragment was identical with rat plzf mRNA in genebank. In the cells 24 h after transfected with recombinant plasmid, Western blot analysis showed a 106 kD fusion protein. Green fluorescence could be seen by fluorescence microscope after 24 h. Conclusion:The recombinant plasmid was successfully constructed and was a useful tool to study the proliferation and differention of spermatogonia.