Objective   To construct the enhanced green fluorescent protein (EGFP)-labeled eukaryotic expression recombinant vector of human syntenin (hSyntenin) gene

and observe its expression in CHG-5 cells.  Methods   hSyntenin was amplified from human glioma by RT-PCR．The PCR products were cloned into the eukaryotic expression vector pEGFP-N1 after XhoⅠ and BamHⅠ digestion. Recombinant plasmid pEGFP-N1-hSyntenin was identified by PCR

double enzyme digestion and DNA sequencing

and transfected into CH

G-5 cells with Lipofectamine 2000. After screened with G418，a stably transfected CHG-5 cell line was established. Expression of hSyntenin in CHG-5 cells was identified by immunocytochemistry and immunoblot analysis

respectively． Results   A positive clone was identified by PCR. The fragments were 4.7 kb and 897 bp in length

which were consistent with the carrier and purpose gene respectively by double enzyme digestion. The eukaryotic expression plasmid of pEGFP-N1-hSyntenin was confirmed by DNA sequencing and its stable transfectant into CHG-5 cell line was established．Fluorescence image revealed that the hSyntenin with green fluorescence was expressed in cytoplasm of CHG-5 cells．Immunoblot following SDS-PAGE showed the positive band at 32 ×10

3

.  Conclusion   Recombinant eukaryotic expression vector of pEGFP-N1-hSyntenin

constructed in this study

is stab1y expressed in CHG-5 cell line.