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Received: 2005-06-03

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Journal of Zhejiang University SCIENCE B 2006 Vol.7 No.1 P.13-19


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

Author(s):  Gong Xing-guo, Ji Jing, Xie Jie, Zhou Yuan, Zhang Jun-yan, Zhong Wen-tao

Affiliation(s):  Institute of Biomacromolecule and Enzyme Engineering, School of Life Sciences, Zhejiang University, Hangzhou 310027, China

Corresponding email(s):   gongxg@cls.zju.edu.cn

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

Gong Xing-guo, Ji Jing, Xie Jie, Zhou Yuan, Zhang Jun-yan, Zhong Wen-tao. Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system[J]. Journal of Zhejiang University Science B, 2006, 7(1): 13-19.

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%T Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system
%A Gong Xing-guo
%A Ji Jing
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T1 - Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system
A1 - Gong Xing-guo
A1 - Ji Jing
A1 - Xie Jie
A1 - Zhou Yuan
A1 - Zhang Jun-yan
A1 - Zhong Wen-tao
J0 - Journal of Zhejiang University Science B
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.2006.B0013

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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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