The DOI matches its Crossref record; 17 of the 17 references it cites were found in a registry or an index.
Tuning Escherichia coli for membrane protein overexpression
Open accessAbstract
A simple generic method for optimizing membrane protein overexpression in Escherichia coli is still lacking. We have studied the physiological response of the widely used “Walker strains” C41(DE3) and C43(DE3), which are derived from BL21(DE3), to membrane protein overexpression. For unknown reasons, overexpression of many membrane proteins in these strains is hardly toxic, often resulting in high overexpression yields. By using a combination of physiological, proteomic, and genetic techniques we have shown that mutations in the lac UV5 promoter governing expression of T7 RNA polymerase are key to the improved membrane protein overexpression characteristics of the Walker strains. Based on this observation, we have engineered a derivative strain of E. coli BL21(DE3), termed Lemo21(DE3), in which the activity of the T7 RNA polymerase can be precisely controlled by its natural inhibitor T7 lysozyme (T7Lys). Lemo21(DE3) is tunable for membrane protein overexpression and conveniently allows optimizing overexpression of any given membrane protein by using only a single strain rather than a multitude of different strains. The generality and simplicity of our approach make it ideal for high-throughput applications.
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Cites
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Consequences of Membrane Protein Overexpression in Escherichia coli
Wagner, S., Baars, L., Ytterberg, A. J., Klussmeier, A., Wagner, C. S., Nord, O., Nygren, P.-Å., van Wijk, K. J., & de Gier, J.-W. (2007) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m600431-mcp200 · Cited by 381
Self-citationOptimization of membrane protein overexpression and purification using GFP fusions
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Self-citationTomoyasu, T., Mogk, A., Langen, H., Goloubinoff, P., & Bukau, B. (2001) · Molecular Microbiology · doi:10.1046/j.1365-2958.2001.02383.x · Cited by 337
Rationalizing membrane protein overexpression
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Recombinant protein expression in Escherichia coli: advances and challenges
Rosano, G. L., & Ceccarelli, E. A. (2014) · Frontiers in Microbiology · doi:10.3389/fmicb.2014.00172 · Cited by 2,556
New tools for recombinant protein production in Escherichia coli: A 5‐year update
Rosano, G. L., Morales, E. S., & Ceccarelli, E. A. (2019) · Protein Science · doi:10.1002/pro.3668 · Cited by 372
Bhatwa, A., Wang, W., Hassan, Y. I., Abraham, N., Li, X.-Z., & Zhou, T. (2021) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2021.630551 · Cited by 317
Jia, B., & Jeon, C. O. (2016) · Open Biology · doi:10.1098/rsob.160196 · Cited by 295
Omadjela, O., Narahari, A., Strumillo, J., Mélida, H., Mazur, O., Bulone, V., & Zimmer, J. (2013) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1314063110 · Cited by 273
Metadata
- Authors
- Wagner, S., Klepsch, M. M., Schlegel, S., Appel, A., Draheim, R., Tarry, M., Högbom, M., van Wijk, K. J., Slotboom, D. J., Persson, J. O., & de Gier, J.-W.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2008-09-23
- Volume
- 105
- Issue
- 38
- Pages
- 14371-14376
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2100100451
- Cited by
- 450
- References
- 17
- FWCI
- 11.07
- Type
- journal article
- Language
- en
- Open access
- green
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- Wagner, S., Klepsch, M. M., Schlegel, S., Appel, A., Draheim, R., Tarry, M., Högbom, M., van Wijk, K. J., Slotboom, D. J., Persson, J. O., & de Gier, J.-W. (2008). Tuning Escherichia coli for membrane protein overexpression. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0804090105
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