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. 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.
The extract is made from the commonly used Spodoptera frugiperda Sf21 cell line. 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 a...
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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. 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.
The extract is made from the commonly used Spodoptera frugiperda Sf21 cell line. 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 incubated at 28–30°C and 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.
Features - Benefits
- Obtain Data Faster: Protein is expressed in only 4 hours, not days as with cell-based expression.
- Complete System: No requirement to purchase additional reagents.
- Achieve High Protein Yields: Express up to 75μg/ml of protein for multiple applications.
Applications
For more information, see the Protein Expression chapter of the Protocols & Applications Guide.
Patents - Disclaimers
Cat.# L1101, L1102: U.S. Pat. Nos. 5,324,637 and 5,492,817 and other patents.
Cat.# L1061, L1081: Patent Pending.
Cat.# L1061, L1081: European Pat. No. 1685247 and other patents pending.
Cat.# L1101, L1102: The method of recombinant expression of Coleoptera luciferase is covered by U.S. Pat. Nos. 5,583,024, 5,674,713 and 5,700,673.
Cat.# L1101, L1102, L1061, L1081: Ezure, T., Suzuki, T., Higashide, S., Shintani, E., Endo, K., Kobayashi, S., Shikata, M., Ito, M., Tanimizu, K., Nishimura, O. (2006) Cell-free protein synthesis system prepared from insect cells by freeze-thawing. Biotechnol. Prog. 22, 1570–7.
Cat.# L1061, L1081: For research use only. Persons wishing to use this product or its derivatives in other fields of use, including without limitation, commercial sale, diagnostics or therapeutics, should contact Promega Corporation for licensing information.
Cat.# L1061, L1081: Use of this vector for applications outside of the TnT® T7 Insect Cell Extract Protein Expression System may require a license from Shimadzu Corporation. For more information contact: t-direct@shimadzu-biotech.jp
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