Molecular Biology Reports

, Volume 37, Issue 3, pp 1335–1340

GST-tagged mouse estrogen receptor α-transactivation domain fusion protein is specifically degraded during its over-expression in E. coli and purification

Article

DOI: 10.1007/s11033-009-9512-8

Cite this article as:
Thakur, M.K. & Ghosh, S. Mol Biol Rep (2010) 37: 1335. doi:10.1007/s11033-009-9512-8

Abstract

Escherichia coli BL21 (DE3) is commonly used for the overproduction of fusion proteins. Using this system, we recently reported the overproduction of histidine-tagged mouse estrogen receptor (ER) α-ligand binding domain as an intact 30 kD protein and its inhibitory effect on the growth of bacteria. However, when GST-tagged mouse ERα transactivation domain (TAD) was overproduced using this system, it showed no effect on the growth of bacteria but was specifically degraded during its expression and purification. Here we report the expression of 47 kD GST-tagged mouse ERα-TAD protein, which was degraded partially and specifically into 46 and 43 kD fragments. This fusion protein was further degraded into 37, 31, 29 and 26 kD fragments during its purification by affinity chromatography. Such specific degradation of GST-tagged mouse ERα-TAD during its overproduction in E. coli and purification indicates the induction of specific protease and suggests the modification of expression system.

Keywords

Estrogen receptor α Fusion protein Protein purification Overproduction Induction Lysate 

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  1. 1.Biochemistry and Molecular Biology Laboratory, Department of ZoologyBanaras Hindu UniversityVaranasiIndia

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