Expression Of Extracellular Domain Of Human Vascular Endothelial Growth Factor Receptor-2 In Pichia Pastoris

Ojective To study the feasibility of the high yield expression of the extracellular domain with the structure of eukaryotic protein of human vascular endothelial growth factor receptor2 (heVEGFR2) in Pichia pastoris.Method We got the heVEGFR2 DNA from pORF-heVEGFR2 by PCR The recombinant Pichia pastoris secreting expression vector (pPICZaA-heVBGFR2) was constructed and transformed into Pichia pastoris X-33 by electroporation . Positive high yield expression recombinant was characterized by its drug-fast phenotype and methanol induction.Result SDS-PAGE shewed the molecular weight of expressed recombinant protein is 108KD.This protein comes up to 45% of total secreted proteins and its concentration comes up to 80mg/L .The molecular weight of its part of the heVEGFR2 is 106.4KD, approximating the molecular weight (116KD) of the recombinant non-chimeric human sKDR protein expressed by insect cells. Western blot analysis showed this protein can bind specifically to the anti- VEGFR2 McAb.Conclusion The heVEGER2 protein with the structure of eukaryotic protein can be expressed high-yieldly in Pichia pastoris...