The TnT® T7 Insect Cell Extract Protein Expression System is a convenient, quick, single-tube, coupled transcription/translation system for the cell-free expression of proteins using an extract derived from the Spodoptera frugiperda Sf21 cell line. Protein synthesis reactions are initiated by the addition of a DNA template, eliminating the need for the time-consuming process of in vitro RNA synthesis.
All components necessary for the transcription/translation are present in the TnT® T7 ICE Master Mix. To initiate protein synthesis, the only component that must be added is the DNA template. Reactions are complete within 4 hours.
Proteins are expressed from genes cloned downstream of the T7 promoter. Companion vectors have been designed to achieve optimal yield with this system (pF25A and pF25K). They contain untranslated region (UTR) sequences at the 5´ and 3´ ends of the gene coding region to enhance translation efficiency. Using the TnT® T7 Insect Cell Extract Protein Expression System and these vectors, 75μg/ml of functional protein can be produced.
For more information, see the Protein Expression chapter of the Protocols & Applications Guide.
GenBank®/EMBL Accession Numbers and Sequence Information
pF25A ICE T7 Flexi® Vector Accession Number EU754721; pF25A ICE T7 Flexi® Vector Vector sequence text file.
pF25K ICE T7 Flexi® Vector Accession Number EU754722; pF25K ICE T7 Flexi® Vector Vector sequence text file.