Construction and expression of the fusion vector for HA-tagged human RAGE gene.
- Author:
Yu-sheng LI
1
;
Xiao-wei GONG
;
Wei-wei CHENG
;
Jie WEI
;
Yong JIANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Base Sequence; Cell Line; Cloning, Molecular; DNA, Complementary; genetics; Gene Expression; Genetic Vectors; genetics; Hemagglutinins; genetics; metabolism; Humans; Molecular Sequence Data; Plasmids; genetics; Receptor for Advanced Glycation End Products; Receptors, Immunologic; biosynthesis; genetics; isolation & purification; Recombinant Fusion Proteins; biosynthesis; genetics; isolation & purification; Sequence Analysis, DNA
- From: Journal of Southern Medical University 2007;27(7):983-986
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a mammalian expression vector for HA-tagged receptor of advanced glycation end products (RAGE).
METHODSHuman RAGE cDNA codon region was amplified by PCR from human cDNA library and cloned into the pcDNA3 vector following routine procedures. After identification by enzyme digestion, PCR and sequencing, the correct recombinant vector RAGE/pcDNA3 was inserted with HA sequence behind the signal peptide sequence of RAGE. After identification by sequencing, HA-RAGE/pcDNA3 was transfected into HEK293 cells, and its expression was detected by Western blotting.
RESULTSIdentification by enzyme digestion, PCR and sequencing, confirmed the validity of the recombinant vector RAGE/pcDNA3, and HA-RAGE/pcDNA3 was highly expressed in HEK 293 cells.
CONCLUSIONHA-tagged RAGE is successfully constructed and expressed in mammalian cells, which may facilitate functional study of RAGE in cell signal transduction.
