1. Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system.
- Author
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Gong XG, Ji J, Xie J, Zhou Y, Zhang JY, and Zhong WT
- Subjects
- Bacterial Proteins biosynthesis, Bacterial Proteins chemistry, Bacterial Proteins genetics, Bacterial Proteins isolation & purification, Glutathione Transferase genetics, Glutathione Transferase isolation & purification, Oncogene Protein pp60(v-src) genetics, Oncogene Protein pp60(v-src) isolation & purification, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins isolation & purification, Saccharomyces cerevisiae genetics, Glutathione Transferase biosynthesis, Oncogene Protein pp60(v-src) biosynthesis, Oncogene Protein pp60(v-src) chemistry, Protein Engineering methods, Saccharomyces cerevisiae metabolism
- 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.
- Published
- 2006
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