The δ-endotoxin proteins accumulate in Escherichia coli as a protein-DNA complex that can be dissociated by hydrophobic interaction chromatography

Chaturvedi, Ratnesh ; Bhakuni, Vinod ; Tuli, Rakesh (2000) The δ-endotoxin proteins accumulate in Escherichia coli as a protein-DNA complex that can be dissociated by hydrophobic interaction chromatography Protein Expression and Purification, 20 (1). pp. 21-26. ISSN 1046-5928

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Official URL: http://linkinghub.elsevier.com/retrieve/pii/S10465...

Related URL: http://dx.doi.org/10.1006/prep.2000.1270

Abstract

The insecticidal protein CryIAc accumulated to form inclusion bodies in Escherichia coli upon overexpression of the cloned gene. The solubilized inclusion bodies contained the δ-endotoxin in association with DNA fragments of about 25 kb. The protein-DNA complex could be dissociated and the δ-endotoxin purified by hydrophobic interaction chromatography on phenyl-sepharose. The DNA was washed out in the high-salt buffer while the δ-endotoxin was bound to the matrix and was eluted at 4°C by a stepwise decreasing potassium chloride gradient. The DNA-protein complex also contained plasmids harbored by the host strain. The plasmid DNA associated with the complex became competent to transform E. coli only after it was dissociated from the δ-endotoxin. The hydrophobic interaction chromatography provides an efficient method for the purification of DNA-free activated toxin.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Bacillus thuringiensis; Crystal Protein; δ-endotoxin; Hydrophobic Interaction Chromatography; Phenyl-sepharose; Protein-DNA Complex; Transformation
ID Code:21022
Deposited On:20 Nov 2010 09:21
Last Modified:16 Nov 2011 14:51

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