Contains the cellular components necessary for protein synthesis (tRNA, ribosomes, initiation, elongation and termination factors). Wheat Germ Extract preparation include grinding in an extraction buffer followed by centrifugation to remove cell debris and chromatography that removes endogenous amino acids and plant pigments from the extract. The extract is also treated with micrococcal nuclease to destroy endogenous mRNA, thus minimizing background translation.
The extract contains an energy-regenerating system (phosphocreatine/phosphocreatine kinase), spermidine to stimulate the efficiency of chain elongation, magnesium acetate and potassium acetate. Potassium Acetate enhances translation for a wide range of mRNAs. Three Amino Acid Mixtures are provided, which enable use of different radioisotopes.
For more information, see the Protein Expression chapter of the Protocols & Applications Guide.
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Applications
- Expression of eukaryotic messages that do not express well in reticulocyte lysates
- Expression of plant mRNAs
- Expression of mammalian transcription factors