Purification and Identification of the Soluble VEGFR-2 Kinases Expressed in Escherichia coli

Abstract:

To improve the soluble expression of VEGFR-2 tyrosine kinase in Escherichia coli for further study on its biological activity.Different concentrations of IPTG,inducing times and temperatures are used to optimize the expression condition for the soluble recombinant VEGFR-2 tyrosine kinase in Escherichia coli.The recombinant protein is purified by affinity chromatography.Western Blot was used to identify this protein.The recombinant plasmid pQE30-TK encoding VEGFR-2 tyrosine kinase was constructed and soluble VEGFR-2 tyrosine kinase was expressed.SDS-PAGE analysis showed that the molecular mass of VEGFR-2 tyrosine kinase was about 36 000 as expected and Western blot analysis indicated that both antibodies for anti-Flk-1 and anti-His could react against the purified protein.Conclusion: The soluble human VEGFR-2 tyrosine kinase was expresses in Escherichia coli by the recombinant technique.The purified protein showed biological activity.