Expression, purification and refolding of extracellular domain of mouse epidermal growth factor receptor.
- Author:
Bing HU
1
;
Ling TIAN
;
You LU
;
Li YANG
;
Xia ZHAO
;
Jiyan LIU
;
Yuquan WEI
Author Information
1. Key Laboratory of Biotherapy of Human Diseases, Ministry of National Education, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041.
- Publication Type:Journal Article
- MeSH:
Animals;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
Mice;
Plasmids;
Receptor, Epidermal Growth Factor;
biosynthesis;
chemistry;
genetics;
isolation & purification;
Transfection
- From:
Journal of Biomedical Engineering
2003;20(2):264-267
- CountryChina
- Language:Chinese
-
Abstract:
A DNA fragment encoding extracellular domain of mouse epidermal growth factor receptor (EGFR) was obtained by PCR from a previous recombinant plasmid. The DNA fragment was then ligated into prokaryotic expression vector, and expressed in Escherichia Coli. The recombinant protein was purified under denature conditions by affinity chromatography, and refolded with gradient dialysis. The recombinant protein could produce antibodies to recognize extracellular domain and full-length of mouse EGFR, and form homodimer in the presence of EGF detected by western blot analysis. These findings provide evidence that the renatured recombinant extracellular domain of mouse epidermal growth factor receptor is immunogenetic and may be important for further application of this protein in functional and immunological research.
