Expression of recombinant h-FGF21 in periplasmic space of Escherichia coli
10.16438/j.0513-4870.2015-1008
- VernacularTitle:成纤维细胞生长因子21在大肠杆菌细胞周质中的表达
- Author:
Shu-jie LI
1
;
Xian-long YE
2
;
Qiang WU
1
;
Yin-hang YU
1
;
Dan YU
1
;
Gui-ping REN
1
;
De-shan LI
1
Author Information
1. College of Life Science, Northeast Agricultural University, Harbin 150030, China
2. College of Life Science, Henan Normal University, Xinxiang 453007, China
- Publication Type:ORIGINAL ARTICLES
- Keywords:
fibroblast growth factor 21;
signal peptide;
periplasmic expression;
diabetes
- From:
Acta Pharmaceutica Sinica
2016;51(5):732-
- CountryChina
- Language:Chinese
-
Abstract:
Fibroblast growth factor 21 (FGF21) is a novel metabolic regulator of glucose and lipid, which is safe, effective and independent on insulin. FGF21 is considered as a prospective anti-diabetic drug. The aim of this study was to express recombinant h-FGF21 in periplasmic space of Escherichia coli. The pET27b plasmid was used to create the expression vectors of h-FGF21 with a PelB secretion signal. The ph-FGF21 (periplasmic expression of h-FGF21) was successfully expressed in the periplasm of E. coli BL21 (DE3), and soluble ph-FGF21 was isolated by disruption of the outer membrane. After twice of ion exchange chromatography, the purity of ph-FGF21 was above 95% in an analysis with a gray analysis software. The molecular weight of ph-FGF21 was about 20 kDa in SDS-PAGE and Western blotting analysis. The activity of ph-FGF21 and ih-FGF21 (intracellular expression of h-FGF21) was observed in vitro in the glucose uptake assay in HepG2 cells. The activity was observed in type 2 diabetic db/db mice after short or long-term treatments. The results suggest that the ph-FGF21 has a consistent activity with ih-FGF21 in vitro and in vivo.
