[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.


CONCLUSIONS
delta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.
Hu
Zi-You
ZY
Research Center of Clinical Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. ziyouhu2002@yahoo.com.cn
Qi
Song-Tao
ST
Zhang
Xia
X
Cao
Qiong
Q
Wu
Bing-Yi
BY
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D010957:Plasmids; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D017465:Receptors, Opioid, delta / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D018076:DNA, Complementary; D017465:Receptors, Opioid, delta; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins