Cloning, expression and glucose regulation activity of human FGF-21
Received date: 2009-10-19
Revised date: 2010-04-13
Online published: 2010-05-24
Fibroblast growth factor (FGF)-21 is a recently discovered glucose regulator and has potential to become therapeutics for treatment of type 2 diabetes. The aim of this study was to clone and express human FGF-21 gene and characterize its bioactivity for glucose regulation. The hFGF-21 cDNA was cloned from human liver by RT-PCR and subcloned into the pSUMO vector after sequencing confirmation. The recombinant plasmid was transformed into Escherichia coli Rosetta strain. The FGF-21 protein expression was induced by IPTG and purified by Ni-NTA agarose. The FGF-21 product was verified by Western blotting analysis with specific antibody. The bioactivity of the purified protein was examined by glucose uptake assay in 3T3-L1 adipocytes. The cloned hFGF-21 gene consisted of 546 bp, which was in agreement with the published data in GenBank. SDS-PAGE analysis showed that hFGF-21 expressed in the E. coli system was 19.4 kDa in size. The glucose uptake assay in 3T3-L1 adipocytes indicated that the purified hFGF-21 could stimulate glucose uptake in a dose-dependent manner, and glucose transporters (GLUT1) is the functional unit.
Key words： human FGF-21; mature protein; diabetes mellitus; SUMO
HOU Yu-Ting, LI Jin-Na, LIN Gui-Ping, LIU Ming-Yao, SUN Guo-Feng, WANG Wen-Fei, LI De-Shan . Cloning, expression and glucose regulation activity of human FGF-21[J]. Hereditas(Beijing), 2010 , 32(6) : 583 -587 . DOI: 10.3724/SP.J.1005.2010.00583
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