[Expression of quail vascular endothelial growth factor receptor quek-1 extracellular domain 2-4 loop cDNA in Pichia pastoris]

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2008 Feb;25(1):157-60, 167.
[Article in Chinese]

Abstract

The extracellular domain 2-4 loop cDNA of quail vascular endothelial growth factor receptor quek1 (qVEGFR2) was obtained from plasmid carrying qVEGFR2 by PCR. Then it was cloned into expression vector pPICZalphaA of Pichia pastoris. To construct recombinant expression plasmid pPICZalphaA-qVEGFR2, linearized pPICZalphaA-qVEGFR2 with SacI was transformed to electroporated Pichia pastoris GS115. Subsequently, positive clone was selected by PCR and its phenotype was determined. SDSPAGE and Western blot assays of culture medium from a methanol-induced expression strain demonstrated that recombinant qVEGFR2 proteins were expressed and secreted into the culture medium. These results could provide a basis for further researches on tumor protein vaccine as well as for the preparation of tumor protein vaccine with Pichia pastoris.

MeSH terms

  • Animals
  • Cancer Vaccines
  • DNA, Complementary / biosynthesis*
  • DNA, Complementary / genetics
  • Genetic Vectors / genetics
  • Pichia / genetics
  • Pichia / metabolism*
  • Polymerase Chain Reaction
  • Quail
  • Receptors, Neurotransmitter / biosynthesis*
  • Receptors, Neurotransmitter / genetics
  • Recombinant Proteins / analysis
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics
  • Vascular Endothelial Growth Factor Receptor-2 / biosynthesis*
  • Vascular Endothelial Growth Factor Receptor-2 / genetics

Substances

  • Cancer Vaccines
  • DNA, Complementary
  • Receptors, Neurotransmitter
  • Recombinant Proteins
  • KDR protein, Coturnix japonica
  • Vascular Endothelial Growth Factor Receptor-2