Tobacco Cell-Free Protein Expression

Tobacco cell-free protein expression technology utilizes tobacco cell-free systems to produce recombinant proteins and allows for efficient mRNA translation in an extracellular environment. Cell-free expression systems are widely used in protein production. Some proteins that are difficult to express and purify in living cells, such as membrane and ion channel proteins, can be produced in cell-free expression systems. Based on an advanced technology platform and rich service experience, Lifeasible provides a high-yield cell-free protein expression system using tobacco cell lysates. Tobacco cell-free protein expression is primarily used to produce antibodies, antigens, enzymes, and membrane proteins. Because it is an animal-free expression system, proteins expressed and purified from the tobacco cell-free system are safer and more suitable for in vivo testing in animals.

Features of Tobacco Cell-Free Protein Expression

• The Tobacco Protein-Free Expression System provides higher expression levels and enables more efficient and scalable protein production than other cell-free protein synthesis systems.
• Tobacco Protein-Free Expression Systems can produce target proteins in a fraction of the time, have shorter production cycles, provide significant time savings in obtaining usable proteins, and facilitate faster testing, optimization, and application development.
• Tobacco protein-free expression systems have the added advantage of enabling post-translational modifications (PTM) typically found in plant-derived proteins. The ability to perform PTMs in a cell-free system that mimics plant processes is beneficial for producing proteins similar to those found in plant systems or proteins that require complex modifications.
• Tobacco protein-free expression systems provide higher purity recovery of expressed proteins, whether they are labeled or not. This is especially beneficial for proteins expressed at low levels or difficult to purify.
• The Tobacco Protein Free Expression System can be used directly in animal experiments for animal source-free proteins, effectively reducing the risk of cross-contamination and addressing the ethical concerns of using animal-source products in research.

Our Service for Tobacco Cell-Free Protein Expression

Subcloning service

We offer services to clone target genes, either PCR products or plasmids, and perform subcloning operations. This includes preparation of target DNA fragments and vectors, ligation, transformation, and recombinant screening.

Transcription reaction services

• Transcription reaction 1. We construct expression vectors containing a T7 or SP6 strong promoter, a tobacco mosaic virus 5' leading sequence containing GAA (translational enhancer), fluorescent proteins, and a multiple cloning site (MCS). After insertion of the target gene into the expression vector, it is transcribed in vitro by interacting with the transcription mixture (including DNA template, SP&RNA polymerase, NTP Mix, Transcription Buffer, and RNase inhibitor).
• Transcription reaction 2. The translation is performed in Translation Buffer (containing 20 amino acids, ATP, and GTP) using the obtained mRNA as a template. We offer translation methods not used in line with the instrument, including the double-layer reaction method, the repetitive batch method, and the filtration flow method, of which the double-layer reaction method is the most widely used.

Product analysis services

We can analyze fluorescent signals, the amount of fluorescent signals, and luciferase activity to ensure the success and activity of protein expression.

Expression optimization

In small-scale screening experiments, we perform meticulous optimization of expression vectors, promoter selection, type and position of affinity purification tags, and optimization of expression conditions to increase yield.

Small-scale purification screening and large-scale protein expression purification

We purify proteins obtained from small-scale experiments by affinity magnetic beads and provide complete lab reports, including SDS-PAGE and WESTERN BLOT data. We can also scale up expression and purification according to the amount required by the customer.

Protein purification

We purify target proteins and can use affinity chromatography, ion exchange chromatography, and gel filtration methods. We can synthesize customized affinity matrices to isolate the active ligand-binding protein fraction as needed.

Protein characterization

We test purified proteins for enzymatic activity, detection of ligand binding, and structural integrity to ensure compliance with relevant requirements.

Fig.2 Our service process. (Lifeasible)

Highlights of Our Tobacco Cell-Free Protein Expression

• Rapid expression services. We offer a tobacco cell-free protein expression system that enables rapid protein expression with a much shorter gene-to-protein time, typically just a few hours.
• High yield. Our systems can achieve high protein expression yields, which can largely help customers who need large quantities of proteins for further research or applications.
• Flexibility and customizability. We can customize expression vectors to meet customer needs, including different promoters, tags, and optimized codons.
• Cost-effectiveness. Compared with a cell culture system, we provide a tobacco cell-free system at a lower cost, which can help customers save money for their projects.

Lifeasible has accumulated rich service experience in microalgae expression system service, and our professional technical team can provide high-quality microalgae expression systems and many related special services. If you are interested, please feel free to contact us.