کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2020552 | 1069188 | 2013 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Characterization and optimization of vascular endothelial growth factor165 (rhVEGF165) expression in Escherichia coli Characterization and optimization of vascular endothelial growth factor165 (rhVEGF165) expression in Escherichia coli](/preview/png/2020552.png)
Vascular endothelial growth factors165 (VEGF165) is the most potent and widely used pro-angiogenic factor. Here we determined optimal culture condition of recombinant human VEGF165 (rhVEGF165) in Escherichia coli (E. coli). rhVEGF165 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, rhVEGF165 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.
► l-Arabinose (0.25% w/v) at 20 °C for 5 h provided optimal culture conditions for rhVEGF165 expression by Escherichia coli.
► Among culture conditions, induction temperature greatly influences the rhVEGF165 expression.
► rhVEGF165 significantly increased cell proliferative activity (p < 0.001) in a dose- and time-dependent manner.
► In addition, rhVEGF165 is involved in endothelial cell growth by upregulation of related genes expression.
Journal: Protein Expression and Purification - Volume 87, Issue 2, February 2013, Pages 55–60