Wheat Germ Extract cell-free expression

Cell-free protein synthesis (CFPS) systems have been widely used for decades as a fast and effective tool in basic biology. Without the need for cell viability and growth, the CFPS system has significant advantages over in vivo protein production systems. Recently, Profacgen has made great efforts to further optimize the CFPS systems, such as E. coli cell-free expression system, wheat germ cell-free expression system, rabbit reticulocyte lysate cell-free expression system, and tobacco cell-free expression system to produce proteins at high yields, reduce costs and expand scale, including simplifying the extract preparation process, developing new energy regeneration systems for protein synthesis, stabilizing substrate supply and promoting the folding of proteins.

Figure 1. Schematic illustration of industrial production of protein therapeutics using CFPS systems. [1]

To maximize the system's translation yield and throughput, Profacgen utilizes the TNT Coupled Wheat Germ Extract Systems to integrate the transcription reaction directly into the translation reaction mixture via a single test tube reaction mode. Unlike ordinary wheat germ extract systems, ordinary wheat germ extracts usually use SP6, T3 or T7 RNA polymerase promoters to synthesize RNA in vitro and then perform translation reactions. TNT reduces the loading step and saves time.

Figure 2. Brief illustration of TNT Coupled Wheat Germ Extract cell-free Systems

Protocal of the wheat germ extract cell-free expression system:

1. We have prepared three kinds of amino acid mixtures for you: methionine-depleted mixture, cysteine-depleted mixture or leucine-depleted mixture as the mother liquor of the reaction components, including tRNA, ribosome, and initiation factor , Elongation factor, termination factor, and with T7, T3 or SP6 RNA polymerase.
2. Add plasmid DNA or PCR products, synthesize mRNA in vitro under the action of RNA polymerase, incubate and detect;
3. Use transcription factors in wheat germ extracts, enzymes required for various synthetic proteins, and supplementary amino acids, energy substances, tRNA, etc. to translate mRNA into proteins and purify the target protein.
4. The mRNA released after translation is recycled using a cell-free system to synthesize the protein, and the mRNA is inactivated after repeated several times.

Advantages of the wheat germ cell-free extract expression system:

1. Save time and increase overall yield of functional, soluble, full-length proteins;
2. Suitable for the expression of toxic proteins such as kinases;
3. Can be labeled with modified tRNA;
4. Can incorporate unnatural amino acids at a specific site;
5. The system can also be used for high-throughput experiments.

Profacgen's cell-free expression platform can provide you with comprehensive technical services, specializing in the special properties of the target protein, and professionally solve protein expression related problems, such as low protein production, expression of special proteins (such as membrane proteins, toxic proteins, etc.), and preparation of proteins Complex, parallel synthesis of many different proteins, etc. We always put your needs first, and if you have any questions about the results, we can help you verify. Profacgen is your ideal partner. If you are interested in our services, please feel free to contact us for more information.

Reference:

[1] Xiaoge Jia, Zixin Deng, Tiangang Liu. Progress of cell-free protein synthesis system and its applications in pharmaceutical engineering - A review[J]. Acta Microbiologica Sinica, 2016, 56(3): 530-542.