Expression and Purification of Receptor Tyrosine Kinase ErbB2 Kinase Domain
- VernacularTitle:ErbB2受体酪氨酸激酶激酶区的表达与纯化
- Author:
Xi JIANG
;
Xiang-Shan ZHOU
;
Yuan-Xing ZHANG
;
- Publication Type:Journal Article
- Keywords:
Receptor tyrosine kinase, Pichia pastoris, Cloning, Expression, RTK activity
- From:
Microbiology
2008;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
The kinase domain of receptor tyrosine kinase(RTK) ErbB2 was expressed fused with GFP in Pichia pastoris. Recombinant expression vector pPIC3.5K was constructed in Escherichia coli TOP10. The right P. pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of recombinant vector, and then induced by methanol in baffled shake bottles. The strain with highest protein yield was scaled up in a 5 L fermentor. Recombinant protein was analyzed with tyrosine kinase assay after Ni2+ affinity chromatograph. Results showed that the 100 kD recombinant protein with tyrosine kinase activity was successfully expressed in P. pastoris.
