Biomolecular Breadboards:DNA parts: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
|||
Line 12: | Line 12: | ||
| | | | ||
* Control sequence: [[Media:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb|pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb]] (GenBank file) | * Control sequence: [[Media:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb|pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb]] (GenBank file) | ||
|} | |} | ||
Revision as of 21:10, 16 July 2012
Home | Protocols | DNA parts | Preliminary Data | Models | More Info |
This page contains a description of the DNA parts used for the biomolecular breadboards.
Control Plasmid: pBEST-OR2-OR1-Pr-UTR1-deGFP-T500
This section contains a description of the deGFP control plasmid for the cell-free expression breadboard.
|
Plasmid description
P70 | Lambda phage promoter OR2-OR1-Pr specific to E. coli σ70 | GenBank: J02459.1 |
UTR1 | The untranslated region containing the T7 g10 leader sequence for highly efficient translation initiation | GenBank: M35614.1 |
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. |
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. |
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.
|
Figure 1. eGFP expression as a function of plasmid DNA template. Plasmid DNA pBEST-OR2-OR1-Pr-UTR1-deGFP-T500 is varied by concentration.