There is no single solution for the expression of all classes of difficult proteins. Instead, expression problem-specific solutions that aim to better the chances of success can be used. For microbial expression systems, these solutions often come in the form of unique host strains that have been genetically modified to enhance the production of a certain difficult protein class. Other expression solutions seek to address problems by controlling aspects of how a target protein is produced. For example, some expression hosts allow for precise control of target gene expression. In addition, certain protein tags can help a protein to more efficiently insert into a host membrane or improve the solubility of a target protein.
- What are the strain properties of Lemo21(DE3) competent E. coli?
- Where can I find more detailed FAQs for the K. lactis Protein Expression Kit?
- What systems does NEB offer for protein expression and purification?
- Which SHuffle® strain should I use?
- How do SHuffle® strains aid in cytoplasmic disulfide bond formation?
- What is LysY?
- What applications are SHuffle® strains useful for?
- High Efficiency Transformation Protocol
- Protein Expression with T7 Express strains
- Expression Using SHuffle (C3030)
- 5 Minute Transformation Protocol (C3026)
- High Efficiency Transformation Protocol
- 5 Minute Transformation Protocol (C3030)
- 5 Minute Transformation Protocol (C3029)
- Expression Using SHuffle (C3026)
- Expression Using SHuffle (C3029)
- Transformation of SHuffle® Competent Cell Strains
- E coli Lemo21 DE3 A T7 RNA Polymerase-based protein overexpression platform for routine and difficult targets
- Protein Expression with T7 Express Strains
- Use of the PURExpress® in vitro Protein Synthesis Kit, Disulfide Bond Enhancer and SHuffle® Competent E. coli for heterologous in vitro and in vivo cellulase expression.
- Using the PURExpress® In Vitro Protein Synthesis Kit for Heterologous In Vitro Expression and Functional Screening of FMN-dependent Oxidoreductase Variants
- NEBExpress® Cell-free E. coli Protein Synthesis System
Avoid Common Obstacles in Protein Expression
Read how to avoid common obstacles in protein expression that prevent interactions with cellular machinery.
The Future of Cell-Free Protein Synthesis
Cell-free protein synthesis has the potential to become one of the most important high throughput technologies for functional genomics and proteomics.
Over 40 years in protein expression and purification – a historical perspective
This article provides an overview of the advances in protein expression and purification methodology over the past 40 years.
- Competent Cell Brochure
- Protein Expression & Purification Brochure
- Competent Cell Product Comparison
- Protein Expression and Purification Selection Chart
- Agrawal, A., Bisharyan, Y., Papoyan, A, Bednenko, J., Cardarelli, J., Yao, M., Clark, T., Berkmen, M., Ke, N., Colussi, P. (2019) Fusion to Tetrahymena thermophila granule lattice protein 1 confers solubility to sexual stage malaria antigens in Escherichia coli. Protein Expr Purif; 153, 7-17. PubMedID: 30081196, DOI: 10.1016/j.pep.2018.08.001.
- Manta, Bruno; Berkmen, Mehmet; (2019) Disulfide Bond Formation in the Periplasm of Escherichia coli. EcoSal Plus; PubMedID: 30761987, DOI: 10.1128/ecosalplus.ESP-0012-2018.
- Leith, E.M., O'Dell, W.B., Ke, N., McClung, C., Berkmen, M., Bergonzo, C., Brinson, R.G., Kelman, Z (2019) Characterization of the internal translation initiation region in monoclonal antibodies expressed in Escherichia coli J Biol Chem; 294(48), 18046-18056.. PubMedID: 31604819, DOI: 10.1074/jbc.RA119.011008
- Reddy, P.T., Brinson, R.G., Hoopes, J.T., McClung, C., Ke, N., Kashi, L. (2018) Platform development for expression and purification of stable isotope labeled monoclonal antibodies in Escherichia coli. mAbs MAbs; 10 (7), 992-1002. PubMedID: 30060704, DOI: 10.1080/19420862.2018.1496879
- Ke, Na; Berkmen, Mehmet; Ren, Guoping; (2017) A water-soluble DsbB variant that catalyzes disulfide-bond formation in vivo Nat Chem Biol; 13, 1022-1028. PubMedID: 28628094, DOI: 10.1038/nchembio.2409
- Ren, G., Ke, N. and Berkmen, M. (2016) Use of the Shuffle Strains in Production of Proteins. Curr Protoc Protein Sci; Aug 1, 1;85:5.26.1-5.26.21.. PubMedID: 27479507 , DOI: 10.1002/cpps.11.
- Anton, B.P., Fomenkov, A., Raleigh, E.A. and Berkmen, M. (2016) Complete Genome Sequence of the Engineered Escherichia coli SHuffle Strains and Their Wild-Type Parents Genome Announc; Mar 31;4(2), PubMedID: 27034504, DOI: 10.1128/genomeA.00230-16.
- Robinson, M.-P., Ke, N., Lobstein, J., Peterson, C., Szkodny, A., Mansell, T.J., Tuckey, C., Riggs, P.D., Colussi, P.A., Noren, C.J., Taron, C.H., Delisa, M.P., Berkmen, M. (2015) Efficient expression of full-length antibodies in the cytoplasm of engineered bacteria Nat Commun; (6)8072, PubMedID: 26311203, DOI: 10.1038/ncomms9072.
- Chatelle C, Kraemer S, Ren G, Chmura H, Marechal N, Boyd D, Roggemans C, Ke N, Riggs P, Bardwell J, Berkmen M (2015) Converting a Sulfenic Acid Reductase into a Disulfide Bond Isomerase Antioxid Redox Signal; 26191605. PubMedID: 26191605, DOI: 10.1089/ars.2014.6235
- Berkmen, M. (2012) Production of disulfide-bonded proteins in Escherichia coli Protein Expr Purif; 240-251. PubMedID: 22085722
- Shouldice, S.R., Cho, S.H., Boyd, D., Heras, B., Eser, M., Beckwith, J., Riggs, P., Martin, J.L.and Berkmen, M. (2010) In vivo oxidative protein folding can be facilitated by oxidation-reduction cycling. Mol Microbiol; 75(1), 13-28. PubMedID: 19968787
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