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[Construction of delta-pIRES2-EGFP plasmid and its expression in HEK293 cells]
Zi-You Hu1, Song-Tao Qi, Xia Zhang
1Research Center of Clinical Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. ziyouhu2002@yahoo.com.cn
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.
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