Biomolecular Breadboards:pBEST OR2 OR1 Pr UTR-deGFP-T500.gb: Difference between revisions
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| The enhanced green fluorescent protein truncated and modified in N- and C- terminal | | The enhanced green fluorescent protein truncated and modified in N- and C- terminal | ||
| Shin, J., and Noireaux, V. (2010) Efficient cell-free expression with the endogenous E. Coli RNA polymerase and sigma factor 70, J Biol Eng 4, 8. | | Shin, J., and Noireaux, V. (2010) Efficient cell-free expression with the endogenous E. Coli RNA polymerase and sigma factor 70, J Biol Eng 4, 8. | ||
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| P70 | |||
| Lambda phage promoter OR2-OR1-Pr specific to E. coli σ70 | |||
| GenBank: J02459.1 | |||
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| T500 | | T500 |
Revision as of 20:46, 16 July 2012
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This page contains a description of the deGFP control plasmid for the cell-free expression breadboard.
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Plasmid description
deGFP | The enhanced green fluorescent protein truncated and modified in N- and C- terminal | Shin, J., and Noireaux, V. (2010) Efficient cell-free expression with the endogenous E. Coli RNA polymerase and sigma factor 70, J Biol Eng 4, 8. |
P70 | Lambda phage promoter OR2-OR1-Pr specific to E. coli σ70 | GenBank: J02459.1 |
T500 | Transcription terminator for E. coli RNA polymerase | Larson, M. H., Greenleaf, W. J., Landick, R., and Block, S. M. (2008) Applied force reveals mechanistic and energetic details of transcription termination, Cell 132, 971-982. |
UTR1 | The untranslated region containing the T7 g10 leader sequence for highly efficient translation initiation | GenBank: M35614.1 |
Preliminary data
Using pBEST-OR2-OR1-Pr-UTR1-deGFP-T500, a plasmid enhanced for GFP expression, the biomolecular breadboard is able to express mass at equal concentrations to comparable bacteriophage in-vitro systems (J. Shin and V. Noireaux, 2010).
Expression of plasmids can be optimized by concentration.
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Figure 1. eGFP expression as a function of plasmid DNA template. Plasmid DNA pBEST-OR2-OR1-Pr-UTR1-deGFP-T500 is varied by concentration.