Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system.
- Author:
Xing-Guo GONG
1
;
Jing JI
;
Jie XIE
;
Yuan ZHOU
;
Jun-Yan ZHANG
;
Wen-Tao ZHONG
Author Information
1. Institute of Biomacromolecule and Enzyme Engineering, School of Life Sciences, Zhejiang University, Hangzhou 310027, China. gongxg@cls.zju.edu.cn
- Publication Type:Journal Article
- MeSH:
Bacterial Proteins;
biosynthesis;
chemistry;
genetics;
isolation & purification;
Glutathione Transferase;
biosynthesis;
genetics;
isolation & purification;
Oncogene Protein pp60(v-src);
biosynthesis;
chemistry;
genetics;
isolation & purification;
Protein Engineering;
methods;
Recombinant Fusion Proteins;
biosynthesis;
chemistry;
isolation & purification;
Saccharomyces cerevisiae;
genetics;
metabolism
- From:
Journal of Zhejiang University. Science. B
2006;7(1):13-19
- CountryChina
- Language:English
-
Abstract:
v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present here the expression, purification, and bioactivity of a GST (glutathione S-transferase)-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl beta-D-thiogalactopyranoside (IPTG)-regulated expression, and the fused protein was purified using GSH (glutathione) affinity chromatography. ELISA (enzyme-linked immunosorbent assay) was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems employed in earlier Src expression.
