Journal of Zhejiang University SCIENCE B

, Volume 7, Issue 1, pp 13–19 | Cite as

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

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 

Document code

CLC number

Q556+.5 Q786 

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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

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