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  • Cell-Free Expression

    Protein Expression

    A rapid method for gene expression analysis, PURExpress® is a novel cell-free transcription/translation system reconstituted from the purified components necessary for E. coli translation.

    PURExpress® is a registered trademark of New England Biolabs, Inc.

    Solutions for the Expression of Difficult Proteins

    NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.

    NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.

    The PURExpress® System Kit Components

    • His-Tagged Translation Factors
      • Initiation Factors (IF1, IF2, IF3)
      • Elongation Factors (EF-Tu, EF-Ts, EF-G)
      • Release Factors (RF1, RF2, RF3)
      • Ribosome Recycling Factor
      • 20 Aminoacyl tRNA synthetases
      • Methionyl tRNA formyltransferase
    • E. coli Ribosomes
    • E. coli tRNAs
    • Energy Regeneration System
    • NTPs, Amino Acids, Salts, Buffer

    In addition, recombinant T7 RNA polymerase is used to couple transcription to translation. The PURE system represents an important step towards a totally defined in vitro transcription/translation system, thus avoiding the “black box” nature of the cell extract-based systems.

    Protein Expression Using the PURExpress® In Vitro Protein Synthesis Kit

    25 µl reactions containing 250 ng template DNA were incubated at 37°C for 2 hours. 2.5 µl of each reaction was analyzed by SDS-PAGE using a 10–20% Tris-glycine gel. Note that proteins can be purified using reverse affinity chromatography (reagents not supplied). The red dot indicates the protein of interest. Marker M is the Protein Ladder (NEB #P7703)