[Construction and expression of the fusion vector for HA-tagged human RAGE gene].
OBJECTIVE
To construct a mammalian expression vector for HA-tagged receptor of advanced glycation end products (RAGE).


METHODS
Human 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.


RESULTS
Identification 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.


CONCLUSIONS
HA-tagged RAGE is successfully constructed and expressed in mammalian cells, which may facilitate functional study of RAGE in cell signal transduction.
Department of Pathophysiology and Key Laboratory of Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515, China.
Yu-sheng
YS
Li
Xiao-wei
XW
Gong
Wei-wei
WW
Cheng
Jie
J
Wei
Yong
Y
Jiang
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006388:Hemagglutinins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D006388:Hemagglutinins; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins