MYtxtl is a fast and easy-to-use kit for in vitro proteins expression. It is a powerful E. coli based cell-free platform combining in vitro transcription (TX) and translation (TL) in a single reaction (TX-TL) enabling one-step protein expression from a DNA template. Our cell-free protein expression kit is based on the TX-TL technology developed at the University of Minnesota.1
Highly flexible protein expression systemMYtxtl relies on the endogenous E. coli transcription and translation machinery, expanding the repertoire of transcription to hundreds of parts2 compared to T7-based in vitro protein expression kits, while still maintaining backward compatibility with T7 promoter driven expression plasmids. As a consequence, this platform has a unique versatility for prototyping biomolecular networks, as proven by the numerous gene circuits that have been designed with it.3 It also correctly executes complex multi-genes molecular programs such as creating infectious phage particles from genomic DNA template.4
- Simple: Just mix your DNA template with our ready-mix containing all necessary component for in vitro protein expression.
- Fast: No need for bacterial transformation, clone selection, cell lysis. Get your expressed protein same day!
- Flexible: Can use plasmid DNA, linear DNA (PCR product) or RNA templates.
- Rapid prototyping: PCR-assemble genes from individual parts (promoter, ORF, terminator) and express them in one day!
- Synthetic Biology: Can express single or multiple proteins at a time for complex circuitry expression.
- Highly tunable: Both RNA and protein turnover can be fine tuned, enabling gene circuitry regulation.
- High yield: Up to 2 mg/ml of expressed protein in batch mode.
Fields of application
- High-throughput analysis of protein-encoding DNA libraries.
- Small scale protein production for screening of protein mutants or ligand libraries.
- Large scale protein production for crystallization (incorporation of Selenomethionine).
- Production of difficult-to-express proteins (Cysteine-rich, toxic).
- Production of functional membrane proteins.
- Direct protein functionalization by incorporation of non-natural, modified or labeled amino acids.
Master Mix Kits - Sigma 70
These kits contain all TX-TL reagents (E. coli extract, energy buffer and amino acids buffer) necessary for coupled transcription - translation as a convenient single tube master mix. The transcription is based on the endogenous core RNA polymerase and the primary sigma factor σ70 present in the E. coli cytoplasmic extract. Therefore all genes to be expressed should be cloned under the control of a σ70 specific promoters. Similarly, circuitry should start with a σ70 specific promoter. Expression of genes under the control of T7 promoter is possible by co-expressing the T7 RNA polymerase under the control of a σ70 specific promoter (plasmid available upon request).
24 Reactions - Master Mix Kit
Catalog number: MYtxtl-70-24-M
This 24 reactions kit contains 3 tubes (75 µl each) of pre-mixed TX-TL reagents (E. coli extract, energy buffer, and amino acids buffer). These reagents are sufficient to prepare 24 x 12 µl TX-TL reactions by mixing 9 µl of MYtxtl Master Mix with 4 µl of DNA template. Kit also contains one tube of positive control template DNA coding for the fluorescent eGFP.
96 Reactions - Master Mix Kit
Catalog number: MYtxtl-70-96-M
This 96 reactions kit contains 12 tubes (75 µl each) of pre-mixed TX-TL reagents (E. coli extract, energy buffer, and amino acids buffer). These reagents are sufficient to prepare 96 x 12 µl TX-TL reactions by mixing 9 µl of MYtxtl Master Mix with 4 µl of DNA template. Kit also contains one tube of positive control template DNA coding for the fluorescent eGFP.
Synthetic DNA templates for expression in TX-TL reactions
We offer a full service of gene synthesis to cover your needs for DNA templates to use in TX-TL reactions. Whether you need plasmid-borne templates or linear fragments of any size up to 10+ Kb, we can deliver. We have also developed a proprietary technology to stabilize linear DNA templates in TX-TL reactions, yielding 10-fold more proteins compared to standard PCR-generated linear templates. Please contact us for your DNA synthesis needs.
Please contact us for pricing.
MYtxtl ManualMYtxtl Current Manual (v01), 2016
1 Shin and Noireaux. Efficient cell-free expression with the endogenous E. coli RNA polymerase and sigma factor 70. Journal of Biological Engineering 4, 8 (2010).
2 Shin and Noireaux. An E. coli Cell-Free Expression Toolbox: Application to Synthetic Gene Circuits and Artificial Cells. ACS Synthetic Biology 1, 29-41 (2012).
3Takahashi et al. Rapidly characterizing the fast dynamics of RNA genetic circuitry with cell-free transcription-translation (TX-TL) systems. ACS Synthetic Biology 4(5), 503-515 (2015).
Siegal-Gaskins et al. Resource usage and gene circuit performance characterization in a cell-free ‘breadboard‘. ACS Synthetic Biology 3(6), 416-425 (2014).
4 Shin et al. Genome Replication, Synthesis, and Assembly of the Bacteriophage T7 in a Single Cell-Free Reaction. ACS Synthetic Biology 1, 408-413 (2012).
5 Shin and Noireaux. Study of messenger RNA inactivation and protein degradation in an Escherichia coli cell-free expression system. J Biol Eng 4, 9 (2010).