[Construction and expression of eukaryotic expression vector for pEGFP-GPC3].

Aim: To construct and expression of eukaryotic expression vector of glypican-3 (GPC3) pEGFP-GPC3 in mouse dendritic cells (DCs).

Methods: The target gene fragment of GPC3 and empty vector were obtained from plasmid pCMV-SPORT6-GPC3 and vector pEGFP-N1 digested by double enzymes. GPC3 and empty vector were linked by ligase T4, and then transfected into E.coli DH5α. Identification with EcoRI, BamHI digestion, the recombinant eukaryotic expression vector pEGFP-GPC3 was transfected into DCs by Lipofectamine(TM);2000. Fluorescent microscopy and Western blot were used to detected the expression of GPC3.

Results: The expression of EGFP-GPC3 fusion protein was detected in DCs, and GPC3 fusion protein of 67 kDa was detected by Western blotting.

Conclusion: The eukaryotic expression vector pEGFP-of GPC3 was constructed succesfully and transfected effectively into DCs.