[Construction of delta-pIRES2-EGFP plasmid and its expression in HEK293 cells].

OBJECTIVE: To construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells. METHODS: Full length cDNA of rat delta opioid receptor gene amplified from rat brain tissues using reverse transcription and nested PCR was cloned into pMD20 T vector. The delta cDNA was inserted into pIRES2-EGFP plasmid to construct the recombinant eukaryotic plasmid delta-pIRES2-EGFP, which was transfected into HEK293 cells via Lipofectamine2000. The expression of delta was examined under fluorescence microscope. RESULTS: The recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid. CONCLUSION: delta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.