Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system
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 s...
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Veröffentlicht in: | Journal of Zhejiang University. B. Science 2006, Vol.7 (1), p.13-19 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | 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 ,B-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. |
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ISSN: | 1673-1581 1862-1783 |
DOI: | 10.1631/jzus.2006.B0013 |