Construction of delta-pIRES2-EGFP plasmid and its expression in HEK293 cells.
- Author:
Zi-You HU
1
;
Song-Tao QI
;
Xia ZHANG
;
Qiong CAO
;
Bing-Yi WU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; DNA, Complementary; genetics; Gene Expression; Genetic Vectors; Green Fluorescent Proteins; biosynthesis; genetics; HEK293 Cells; Humans; Plasmids; Polymerase Chain Reaction; Rats; Rats, Sprague-Dawley; Receptors, Opioid, delta; genetics; Recombinant Fusion Proteins; biosynthesis; genetics; Transfection
- From: Journal of Southern Medical University 2009;29(7):1351-1353
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells.
METHODSFull 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.
RESULTSThe recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid.
CONCLUSIONdelta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.
