Recombinant production of proteins is one of the most powerful techniques used in the Life Sciences. The ability to produce and purify an abundance of a desired recombinant protein can permit a wide range of possibilities including, its use in industrial processes, or its use to diagnose or treat disease.
At first glance, recombinant protein expression looks quite simple. Essentially, DNA encoding a target protein is cloned downstream of a promoter in an expression vector. This vector is then introduced into a host cell, and the cell’s protein synthesis machinery produces the desired protein. In practice, however, protein expression can be very challenging because so many factors may influence the process. For example, each protein folds in its own unique manner, a process that may be influenced by the choice of expression host. Similarly, some proteins require post-translational modifications or proper insertion into a biological membrane. Finally, some proteins may have an activity that is detrimental to the host. Thus, no single solution exists for successful production of all recombinant proteins. Instead, it is beneficial to have access to a wide range of expression tools, and a willingness to explore multiple approaches to better one’s chances for success.
Protein Expression & Purification includes these areas of focus:
- Protein Expression Approaches
- E.coli Protein Expression
- Yeast Protein Expression
- Cell-Free Protein Expression
- Coupled Protein Expression & Purification
- Maltose Binding Protein Expression
- IMPACT™ Protein Expression
- His-tagged Protein Expression
- Yeast Chitin Binding Domain Tag
- Expression of Difficult Proteins
- Toxic Protein Expression
- Membrane Protein Expression
- Disulfide-Bonded Protein Expression
- Affinity Purification
FAQs for Protein Expression & Purification
Protocols for Protein Expression & Purification
Application Notes Protein Expression & Purification
- E. coli Lemo21(DE3)
- Co-expression of Multiple Proteins in Kluyveromyces lactis
- Protein Expression with T7 Express Strains
- Transformation of SHuffle® Competent Cell 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
Bypassing Common Obstacles in Protein Expression
The Next Generation of Cell-free Protein Synthesis
Why Choose the K. lactis Protein Expression Kit?
Review the advantages of the K. lactis Protein Expression Kit for rapid, high yield protein expression in yeast.
Competent Cells Brochure
The Competent Cells brochure provides information on the different competent cell strains for cloning and protein expression available from NEB.
Protein Expression & Purification Brochure
The Protein Expression and Purification brochure provides information on the advanced tools for protein expression and purification offered by NEB.
- Development of a Rapid Method for Genome Engineering in a Quest Towards Customized Protein Expression Strains (2015)
- NiCo21(DE3): A BL21(DE3) Derivative Designed for Expression and Purification of His-tagged Recombinant Protein (2015)
- Competent Cell Product Comparison
- Protein Expression and Purification Selection Chart
- Convenient Formats of Competent Cells
Other Tools & Resources
NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.