Cell-free Protein Expression
- Expression Vectors for in vitro Protein Synthesis
- in vitro Transcription and Translation (TXTL) Single Tube Protein Synthesis Technology
• Only standard laboratory equipment required
• Fast Processing - no transformation, clone selection and cell lysis
• Different optimized kits for specific needs
• Compatible with E. coli-specific and T7 expression systems
• Non-GMO system with laboratory biosafety level
myTXTL® is a a comprehensive
solution for protein engineering
and synthetic biology applications
myTXTL® is a versatile and easy-to-use cell-free protein expression platform for protein synthesis and synthetic biology in industry and academia. Originally developed by Vincent Noireaux, PhD at the University of Minnesota, this revolutionary and powerful TXTL technology was further refined at Arbor Biosciences toward even greater robustness and to deliver reliable and consistent performance.
In all myTXTL® kits, gene transcription (TX) and translation (TL) is executed in a single reaction tube utilizing the endogenous TXTL machinery from E. coli. Gene expression is initialized by simply adding the nucleotide template to the myTXTL® Master Mix and recombinant proteins can be detected after only a few minutes. Due to the lack of cell walls and the independency from cell growth and viability, cell-free systems allow rapid and parallel production of soluble or toxic proteins and other complex biological structures under open-reaction conditions. This enables facile manipulation and straightforward downstream processing of synthesized material.
The myTXTL® technology is well-characterized and has been employed for various applications in protein engineering and synthetic biology applications. The myTXTL® Toolbox 2.0 Plasmid Collection contains over one hundred plasmids with various promoters and open reading frames (ORFs) to investigate gene regulation and molecule turnover. Available open reading frames include a wide selection of transcription factors, TXTL modulators and fluorescent reporter proteins to build multi-stage gene circuits.
myTXTL® unites an extremely user-friendly and safe, GMO-free, handling with exceptional protein synthesis efficiency and great versatility, which makes it similarly well-suited for cutting-edge research and student education.
• Protein Expression
• Synthetic Biology
• HTS Protein Engineering
• Protein Evolution
• Gene Circuits & Networks
• TXTL with Linear DNA Templates
• Membrane Proteins
• Protein Functionalization
• Molecular Interaction Analysis
Figure 1. Workflow
After template DNA – either circular or linear – has been added to the ready-to-use Sigma 70 Master Mix, in vitro protein expression starts immediately. The open-reaction environment allows easy manipulation of the system and straight-forward downstream processing of the recombinant protein.
Figure 2. Effect of plasmid concentration on in vitro protein production.
deGFP expression is regulated by the interaction of the endogenous E. coli core RNA polymerase and the primary sigma factor ó70 with the ó70-specific promoter P70a.
Figure 3. Example of a two-plasmid gene network.
Gene expression under the control of the bacteriophage T7 promoter/operator system facilitated by initial expression of T7 RNA polymerase.
Garenne, D. et al. (2018) Cell-free transcription–translation: engineering biology from the nanometer to the millimeter scale. Current Opinion in Biotechnology 58: 19-27.
Marshall, R. et al. (2018) Rapid and scalable characterization of CRISPR technologies using an E. coli cell-free transcription-translation system. Molecular Cell 69: 146-157.
Rustad M. et al. (2018) Cell-free TXTL synthesis of infectious bacteriophage T4 in a single test tube reaction. Synthetic Biology.
Garamella J. et al. (2016) The all E. coli TX-TL toolbox 2.0: a platform for cell-free synthetic biology. ACS Synth Biol. 5:344-355.
Caschera F. et al. (2014) Synthesis of 2.3 mg/mL of protein with an all Escherichia coli cell-free transcription-translation system. Biochimie 99:162-168.