Synthesis of the human VEGF165 gene based on overlap PCR and recombinant expression in stable transfected CHO cells

Abstract

Vascular endothelial growth factors (VEGFs) are a member of a family of structurally related proteins that mediate angiogenesis, and vascular maintenance. VEGF₁₆₅ is an isoform and has been studied extensively because of its potential use in therapeutic angiogenesis. In this study, we synthesized the human VEGF₁₆₅ (hVEGF₁₆₅) gene based on overlap PCR method and recombinant expressed in Chinese’s hamster ovary (CHO) cell. The coding sequence of hVEGF165 was synthesized successfully and the amounts of the recombinant protein could reach to 603.085 pg/ml. This rhVEGF₁₆₅ protein could promote angiogenesis in chicken embryos. In conclusion, we established the stable expression system of recombinant human VEGF₁₆₅ (rhVEGF₁₆₅) protein in CHO cells.

Key words: Overlap extension PCR (OE-PCR), vascular endothelial growth factor (VEGF), Chinese’s hamster ovary (CHO) cell, recombinant protein.