Cell-Free Protein Expression Systems: A Rapid and Efficient Tool for Protein Synthesis

Cell-free protein expression systems are innovative tools that utilize exogenous mRNA or DNA templates to synthesize proteins in vitro, using cell extracts or reconstructed systems containing the necessary enzymes and cofactors. These systems include all the templates, molecular machinery, substrates, and energy required for protein expression, and they are not constrained by cellular metabolism or the enclosed cellular environment. This allows for the rapid and high-yield expression of proteins outside of living cells. Commonly used cell-free protein expression systems include E. coli-based systems, insect cell lysates, wheat germ systems, and rabbit reticulocyte lysates.

Components of Cell-Free Protein Expression Systems

A typical cell-free protein expression system includes several essential components:

• Protein Gene Template: Exogenous mRNA or DNA serving as the blueprint for protein synthesis.
• Energy Source: High-energy molecules like ATP, providing the necessary energy for the reactions.
• Enzymes for Protein Synthesis: Such as RNA polymerase and various translation factors.
• Ribosomes: Molecular machines responsible for synthesizing proteins.
• Amino Acids: The building blocks of proteins.
• Coupled Transcription-Translation System: Simultaneously conducting mRNA synthesis and protein translation.

Advantages of Cell-Free Protein Expression Systems

Cell-free protein expression systems offer several significant advantages:

1. Small Reaction Volume: Capable of synthesizing multiple different proteins in parallel.
2. Incorporation of Unnatural Amino Acids: Enables the inclusion of non-natural or isotope-labeled amino acids for the expression of specialized proteins.
3. Lack of Cellular Structure Constraints: Suitable for producing exogenous proteins that may be toxic to host cells.
4. Short Reaction Time: Meets the needs of high-throughput ligand screening and proteomics studies.
5. High Efficiency: Eliminates steps such as plasmid transformation, cell culture, harvesting, lysis, and centrifugation.
6. Improved Expression of Complex Proteins: Effective for proteins that are difficult to express in traditional cell systems due to multiple transmembrane domains or high hydrophobicity.
7. Open Reaction System: Facilitates the modification of reaction conditions, allowing better control of gene transcription, protein synthesis, and post-translational modifications, while avoiding the formation of inclusion bodies.

E. coli Cell-Free Expression System

The E. coli cell-free protein expression system is the most widely used among cell-free systems. Its features include:

• Clear Genetic Background: Well-understood genetics of E. coli makes gene manipulation straightforward.
• High Expression Levels: Achieves high levels of target protein expression.
• Short Cultivation Cycle: Fast expression cycle and high efficiency.
• Low Cost: Simple operation and low cost.
• Strong Contamination Resistance: High stability and contamination resistance.
• Scalability: Suitable for expressing various proteins.

SBS Genetech's

E. coli Cell-Free Protein Expression Kit

SBS Genetech's E.coli Cell-Free Protein Expression Kit enables the rapid and efficient expression of target proteins using plasmid or linear templates. This kit employs a coupled transcription and translation method to express target proteins through a "one-pot" reaction. The cell-free reaction system provided in the kit includes all necessary enzymes, energy sources, amino acids, nucleotides, and inorganic salts required for in vitro transcription and translation, ensuring simple operation and high expression efficiency.

Product Advantages

1. Speed: Express target proteins in as little as 1 hour.
2. Template Flexibility: Supports linear templates, plasmid templates, and PCR products.
3. High Yield: Achieve protein yields of over 3 mg/mL.
4. Co-Expression: Allows simultaneous expression of multiple target proteins in a single reaction.
5. Simplicity: Single-step reaction—simply mix the reaction components with the DNA template to produce proteins.
6. High Throughput: Reactions can be performed in a 96-well plate, and upon detecting positive expression, the system can be scaled up.
7. Versatility: Capable of expressing various types of proteins, including those rich in disulfide bonds and membrane proteins.

Conclusion

Cell-free protein expression systems serve as a rapid and efficient tool for protein synthesis, with extensive applications in both fundamental research and industrial production. These systems play a critical role in areas such as protein structure and function analysis, high-throughput screening, and more. With continuous optimization and improvement, cell-free protein expression systems are poised to drive further breakthroughs and innovations in the life sciences and biotechnology industries.

SBS Genetech's E.coli Cell-Free Protein Expression Kit exemplifies these advances, providing researchers with a powerful and versatile tool for high-quality protein expression. Our kit offers unmatched speed, flexibility, and efficiency, making it an ideal choice for both academic research and industrial applications. Whether you need to express complex proteins, perform high-throughput screening, or conduct detailed protein studies, our kit is designed to meet your needs.

Why Choose SBS Genetech's

E.coli Cell-Free Protein Expression Kit?

• Unparalleled Speed: Achieve protein expression in as little as 1 hour.
• High Yield: Obtain protein yields exceeding 3 mg/mL.
• Versatility: Suitable for a wide range of protein types, including those with disulfide bonds and membrane proteins.
• Simplicity and Efficiency: One-step reaction process simplifies your workflow.
• Scalability: Easily scale up your experiments from 96-well plates to larger volumes.

We invite you to experience the benefits of our E.coli Cell-Free Protein Expression Kit and see how it can transform your protein expression experiments. Visit our website or contact us directly to place your order and join the growing number of researchers who rely on SBS Genetech for their protein synthesis needs. Let us help you accelerate your research and achieve your scientific goals with ease and efficiency.