Cloning, expression and purification of KDR tyrosine kinase.
- Author:
Chunping LIU
1
;
Yang ZHANG
;
Yuan LI
Author Information
1. Key Laboratory of Biotechnology of Antibiotics, Ministry of Health, Institute of Medicinal Biotechnology, Beijing 100050, China.
- Publication Type:Journal Article
- MeSH:
Catalytic Domain;
genetics;
Cloning, Molecular;
Endothelial Cells;
cytology;
enzymology;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
Humans;
Recombinant Proteins;
biosynthesis;
genetics;
isolation & purification;
Umbilical Veins;
cytology;
Vascular Endothelial Growth Factor Receptor-2;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2008;24(9):1545-1549
- CountryChina
- Language:Chinese
-
Abstract:
The catalytic domain of KDR kinase (KDR-CD) was amplified from RNA of HUVCEs cells with RT-PCR and expressed in E. coli BL21(DE3) by plasmid pET30a as vector. The recombinant protein was purified with affinity chromatography (Ni-NTA). Western blotting showed that the recombinant KDR-CD was phosphorylated in E. coli BL21(DE3). The recombinant KDR-CD was identified to have kinase activity catalyzing the substrate phosphorylated with ATP in the enzymatic reaction.
