title: High-level Expression and Purification of Active Human FGF-2 in Escherichia coli by Codon and Culture Condition Optimization.
abstract: BACKGROUND
  Basic fibroblast growth factor (bFGF) is a member of a highly conserved superfamily of proteins that are involved in cell proliferation, differentiation, and migration.
  
  
  OBJECTIVE
  The objective of this study was to overexpress and purify the high-level active human bFGF in Escherichia coli (E. coli).
  
  
  METHODS
  This experimental study was conducted in the Islamic Republic of Iran. After codon optimization and gene synthesis, the optimized FGF-2 gene was subcloned into plasmid pET-32a. pET32-FGF-2 was transformed into E. coli BL21 for expression. The cultivation parameters were optimized to produce a high yield of FGF-2.
  
  
  RESULTS
  The optimal conditions were determined as follows: cultivation at 37°C in TB medium, with 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG), followed by post-induction expression for 6 h. Under the abovementioned conditions, the expression volumetric productivity of FGF-2 reached 1.48 g/L.
  
  
  CONCLUSIONS
  A fusion tag from the pET32 expression plasmid permits the recovery of the recombinant fusion FGF-2 from E. coli, without affecting its biological activity.
authors.affiliation: Department of Biotechnology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Soleyman
authors.affiliation: Department of Biotechnology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors.forename: Mostafa
authors.initials: M
authors.lastname: Khalili
authors.affiliation: Department of Microbilogy and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors.forename: Behzad
authors.initials: B
authors.lastname: Khansarinejad
authors.affiliation: Department of Anatomy, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Baazm
keywords: Codon; E. coli; Fibroblast Growth Factor 2; Protein