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Updated: Aug 6, 2026
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 26, 2010
[Expression and characterization of human vascular endothelial growth factor in Pichia pastoris]
1Institute of Molecular Immunology, First Military Medical University, Guangzhou 510515, China.
Objective:
To study the expression of human VEGF165 cDNA in Pichia pastoris and to obtain high-level expression of recombinant human VEGP165 (hVEGF165) with good biological activity.
Methods:
Amplifying hVEGF165 cDNA by PCR, after confirmed by DNA sequence analysis, the gene was inserted into the Pichia pastoris expression vector pPIC9K containing AOX1 promoter and a secreting signal peptides, the recombinant expression plasmid pPIC9K/VEGF165 was constructed and transformed into KM71. The multiple insert transformants were screened, fermented in flasks and induced by 1% methanol.
Results:
After 4 days of methanol induction, the expressed hVEGF165 came up to 30% of total proteins in supernatant by SDS-PAGE. The expressed hVEGF165 was further proved having good antigenicity and high specificity by ELISA and Western blot assay, and having good biological activity to stimulate HUVEC proliferation.
Conclusions:
High-level expression of secreted hVEGF165 had been successfully achieved in Pichia pastoris expression system.
