[Efficient expression of soluble human FGF-21 and its glucose regulation activity].
The cDNA of human FGF-21 was subcloned into the pSUMO expression vector and the fusion protein was induced to express in Escherichia coli Rosetta (DE3). The recombinant hFGF-21 was expressed in soluble form in the pSUMO expression system. The recombinant fusion protein was purified by Ni-NTA column. The purified recombinant protein was dialyzed against PBS for re-nature. To obtain pure and active recombinant protein, the fusion protein was subjected to cleavage with SUMO protease I. To examine glucose regulation activity of hFGF-21, 3T3-L1 pre-adipocytes were differentiated into adipocytes, glucose up-take activity of hFGF-21 was examined by glucose oxidase and peroxidase (GOD-POD) assay. Compared with no stimulation control, the recombinant hFGF-21 treatment led to a significant increase in glucose consumption of adipocytes and a significant decrease in concentration of glucose in the medium (P < 0.05, P < 0.001).
Ren
Gui-ping
GP
College of Life Science of Northeast Agricultural University, Provincial Academy of Agricultural Sciences of Heilongjiang Postdoctoral Station, Northeast Forestry University Postdoctoral Station, Harbin 150030, China.
Hou
Yu-ting
YT
Jiang
Yuan-yuan
YY
Li
Jin-nan
JN
Zhang
Wei
W
Di
Liu
L
Li
De-shan
DS
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005346:Fibroblast Growth Factors / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D051379:Mice; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; C414620:fibroblast growth factor 21; D005346:Fibroblast Growth Factors; D005947:Glucose