Multi-protein complexes, critical for cellular processes, rely on individual protein properties and inter-protein interactions. In prokaryotes, heterologous proteins often misfold and lose activity due to insufficient cofactors or post-translational modifications. E. coli co-expression technology simulates natural interaction environments by expressing multiple proteins simultaneously, yielding correctly folded, bioactive complexes. It is widely used in biochemical analysis, structural biology, and high-throughput screening, serving as an efficient, low-cost alternative to yeast or mammalian cell systems.
Biotyscience offers professional E. coli protein co-expression services with advanced platforms and experience. Covering experimental design to protein identification, we provide customized solutions for diverse co-expression needs, ensuring efficient, active, and pure protein complexes to accelerate research.
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Website: www.biotyscience.com Phone: + 400-669-8850 Email: biotyscience@gmail.com
Phase | Procedure | Timeline |
Vector Design and Construction | 1. Select co-expression strategies (single-vector/multi-vector systems); 2. Design combinations of promoters, tags, and replication origins; 3. Perform target gene cloning, vector digestion and ligation, sequencing verification, and integrate tags and regulatory elements | 1-2 weeks |
Host Preparation and Positive Clone Screening | 1. Select suitable strains; 2. Prepare culture media and inducers; 3. Transform plasmids into host cells; 4. Obtain positive clones through resistance screening and PCR verification | 1 weeks |
Optimization of Expression Conditions and Large-Scale Cultivation | 1. Conduct small-scale expression using 24/96-well plates and optimize induction conditions; 2. Scale up cultivation via shake flasks or fermenters | 1 weeks |
Protein Purification and Identification | 1. Lyse cells by ultrasonic treatment/lysozyme, purify through affinity chromatography and remove impurity proteins; 2. Verify products by SDS-PAGE and Western blot, and confirm the function of the complex through activity determination | 1 weeks |
High-Efficiency Expression
Cost-Effective Solution
Guaranteed High Activity
Flexible and Scalable
Reference
Haffke, M., Marek, M., Pelosse, M., Diebold, M. L., Schlattner, U., Berger, I., & Romier, C. (2014). Characterization and production of protein complexes by co-expression in Escherichia coli. In Structural Proteomics: High-Throughput Methods (pp. 63-89). New York, NY: Springer New York.
Chen, H., Huang, R., & Zhang, Y. H. P. (2017). Systematic comparison of co-expression of multiple recombinant thermophilic enzymes in Escherichia coli BL21 (DE3). Applied microbiology and biotechnology, 101(11), 4481-4493.
Busso, D., Peleg, Y., Heidebrecht, T., Romier, C., Jacobovitch, Y., Dantes, A., ... & Celie, P. H. (2011). Expression of protein complexes using multiple Escherichia coli protein co-expression systems: a benchmarking study. Journal of Structural Biology, 175(2), 159-170.
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