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Journal of Zhejiang University SCIENCE B

ISSN 1673-1581(Print), 1862-1783(Online), Monthly

Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system

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 β-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.

Key words: v-Src, GST-fusion, Inclusion body, Orthogonalization, Protein tyrosine kinase


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DOI:

10.1631/jzus.2006.B0013

CLC number:

Q556+.5; Q786

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Received:

2005-06-03

Revision Accepted:

2005-09-09

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