title: Characterization and optimization of vascular endothelial growth factor(165) (rhVEGF(165)) expression in Escherichia coli.
abstract: Vascular endothelial growth factors(165) (VEGF(165)) is the most potent and widely used pro-angiogenic factor. Here we determined optimal culture condition of recombinant human VEGF(165) (rhVEGF(165)) in Escherichia coli (E. coli). rhVEGF(165) expression was the highest in 0.25% of L-arabinose induction concentration, at 20 °C induction temperature, and for 5 h induction time under the control of araBAD promoter using pBADHisA vector. In biological activity test, rhVEGF(165) significantly increased the proliferative activity of CPAE cells (p<0.001) and upregulated the expressions of endothelial cell growth-related genes, such as platelet endothelial cell adhesion molecule (PECAM-1), endothelial-specific receptor tyrosine kinase (TEK), kinase insert domain protein receptor (KDR), and tyrosine kinase with immunoglobulin-like and EGF-like domains 1 (TIE1) in calf pulmonary artery endothelial (CPAE) cells.
authors.affiliation: Department of Biochemistry, Inha University School of Medicine, Incheon 400-712, Republic of Korea.
authors.forename: W
authors.initials: W
authors.lastname: Kang
authors.forename: S
authors.initials: S
authors.lastname: Kim
authors.forename: S
authors.initials: S
authors.lastname: Lee
authors.forename: E
authors.initials: E
authors.lastname: Jeon
authors.forename: Y
authors.initials: Y
authors.lastname: Lee
authors.forename: Y R
authors.initials: YR
authors.lastname: Yun
authors.forename: C K
authors.initials: CK
authors.lastname: Suh
authors.forename: H W
authors.initials: HW
authors.lastname: Kim
authors.forename: J H
authors.initials: JH
authors.lastname: Jang
mesh_terms: D000818:Animals; D001089:Arabinose; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
chemical_list: D003470:Culture Media; D012333:RNA, Messenger; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D001089:Arabinose