Journal article
Characterization of factors favoring the expression of soluble protozoan tubulin proteins in Escherichia coli
Protein Expression and Purification, Vol.29(1), pp.117-122
2003
Abstract
The α- and β-tubulin genes of the parasitic protozoa Giardia duodenalis, Cryptosporidium parvum, and Encephalitozoon intestinalis have been overexpressed in soluble form using Escherichia coli-based expression systems. Several expression systems were compared in terms of the amount of soluble protein produced with different fusion partners, strains of E. coli BL21, and expression temperatures. The cleavability of the fusion partner was also assessed in terms of post-expression applications of the recombinant protein. The maltose-binding protein (MBP) and glutathione S-transferase (GST) fusion partners produced the highest expression levels for all six proteins without the formation of inclusion bodies. The expression system also provided a means of purifying the soluble protein using affinity and anion-exchange chromatography while minimizing protein losses. The yield and purity were therefore very high for both the MBP and GST systems. The tubulin monomers were demonstrated to be assembly-competent using a standard dimerization assay and also retained full antigenicity with monoclonal antibodies. This study presents several methods which are suitable for producing soluble tubulin monomers and, thus, circumventing the formation of inclusion bodies which necessitates re-folding of the tubulin.
Details
- Title
- Characterization of factors favoring the expression of soluble protozoan tubulin proteins in Escherichia coli
- Authors/Creators
- L.M. MacDonald (Author/Creator) - Murdoch UniversityA. Armson (Author/Creator) - Murdoch UniversityR.C.A. Thompson (Author/Creator) - Murdoch UniversityJ.A. Reynoldson (Author/Creator) - Murdoch University
- Publication Details
- Protein Expression and Purification, Vol.29(1), pp.117-122
- Publisher
- Academic Press
- Identifiers
- 991005544331507891
- Copyright
- © 2003 Elsevier Science (USA).
- Murdoch Affiliation
- School of Veterinary and Biomedical Sciences
- Language
- English
- Resource Type
- Journal article
UN Sustainable Development Goals (SDGs)
This output has contributed to the advancement of the following goals:
Source: InCites
Metrics
75 Record Views
InCites Highlights
These are selected metrics from InCites Benchmarking & Analytics tool, related to this output
- Citation topics
- 2 Chemistry
- 2.166 Chromatography & Electrophoresis
- 2.166.873 Protein Purification
- Web Of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- Biotechnology & Applied Microbiology
- ESI research areas
- Biology & Biochemistry