High-level expression of soluble viral structural protein in escherichia coli

Chuan, Y. P., Lua, L. H. L. and Middelberg, A. P. J. (2008) High-level expression of soluble viral structural protein in escherichia coli. Journal of Biotechnology, 134 1-2: 64-71. doi:10.1016/j.jbiotec.2007.12.004


Author Chuan, Y. P.
Lua, L. H. L.
Middelberg, A. P. J.
Title High-level expression of soluble viral structural protein in escherichia coli
Journal name Journal of Biotechnology   Check publisher's open access policy
ISSN 0168-1656
Publication date 2008-03-20
Year available 2008
Sub-type Article (original research)
DOI 10.1016/j.jbiotec.2007.12.004
Volume 134
Issue 1-2
Start page 64
End page 71
Total pages 8
Editor Puehler, A.
Place of publication Netherlands
Publisher Elsevier BV
Collection year 2009
Language eng
Subject C1
100799 Nanotechnology not elsewhere classified
100302 Bioprocessing, Bioproduction and Bioproducts
860899 Human Pharmaceutical Products not elsewhere classified
09 Engineering
0999 Other Engineering
Abstract Pharmaceutically relevant virus-like particles (VLPs) can potentially be manufactured cheaply and efficiently through in vitro assembly of viral structural protein in cell-free reactors, but a bottleneck for this processing route is the currently low-level expression of soluble viral protein in efficient cell factories such as Escherichia coli (E coli). Here, we report expression levels of up to 180 mg L-1 that are achievable from low-cell-density E. coli cultures using a simple and low cost strategy. We investigated effects of host strain, plasmid, inducer concentration, pre-induction temperature and cell density at induction with design of experiment (DOE). The statistical approach successfully identified significant effects and their interactions, and provided insights into the role of codon-usage effects in expression of viral structural protein. In particular, our results support the notion that full codon optimization may be unnecessary to improve expression of viral genes rich in E. coli rare codons; using a strategically modified host cell could provide a simpler and cheaper alternative.
Keyword Codon Optimization
Design of Experiment
Escherichia Coli
Murine Polyomavirus
Soluble Structural Proteins
Virus-Like Particles
Q-Index Code C1
Q-Index Status Confirmed Code

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Fri, 17 Apr 2009, 08:51:59 EST by Amanda Lee on behalf of Aust Institute for Bioengineering & Nanotechnology