Journal of Zhejiang University SCIENCE B

, Volume 7, Issue 1, pp 13–19

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

  • Gong Xing-guo 
  • Ji Jing 
  • Xie Jie 
  • Zhou Yuan 
  • Zhang Jun-yan 
  • Zhong Wen-tao 
Article

DOI: 10.1631/jzus.2006.B0013

Cite this article as:
Gong, X., Ji, J., Xie, J. et al. J. Zhejiang Univ. - Sci. B (2006) 7: 13. doi:10.1631/jzus.2006.B0013

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-SrcGST-fusionInclusion bodyOrthogonalizationProtein tyrosine kinase

Document code

A

CLC number

Q556+.5Q786

Copyright information

© Zhejiang University 2006

Authors and Affiliations

  • Gong Xing-guo 
    • 1
  • Ji Jing 
    • 1
  • Xie Jie 
    • 1
  • Zhou Yuan 
    • 1
  • Zhang Jun-yan 
    • 1
  • Zhong Wen-tao 
    • 1
  1. 1.Institute of Biomacromolecule and Enzyme Engineering, School of Life SciencesZhejiang UniversityHangzhouChina