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| {{Template:Biomolecular Breadboards}}
| | #REDIRECT [[Biomolecular Breadboards:DNA parts]] |
| This page contains a description of the deGFP control plasmid for the cell-free expression breadboard.
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| {|
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| | [[Image:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.png]]
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| * Control sequence: [[Media:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb|pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb]] (GenBank file)
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| * Preliminary data: [[Media:plasmid_deGFP-070812.xls|plasmid_deGFP-070812.xls]] (Excel spreadsheet)
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| |}
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| === Plasmid description ===
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| {| border=1 width=100% cellpadding=4
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| | P70
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| | Lambda phage promoter OR2-OR1-Pr specific to E. coli σ70
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| | GenBank: J02459.1
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| | UTR1
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| | The untranslated region containing the T7 g10 leader sequence for highly efficient translation initiation
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| | GenBank: M35614.1
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| | deGFP
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| | The enhanced green fluorescent protein truncated and modified in N- and C- terminal
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| | 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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| | T500
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| | Transcription terminator for E. coli RNA polymerase
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| | 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.
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| |}
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| === Preliminary data ===
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| 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).
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| Expression of plasmids can be optimized by concentration.
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| {|
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| [[Image:plasmid_sat.png|400px]]
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| * Raw data file: [[Media:plasmid_deGFP-070812.xls|plasmid_deGFP-070812.xls]] (Excel spreadsheet)
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| * Information about protocol on "Outline" sheet; plotted data is on the sheet "plotted-data"
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| <hr>
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| * Control plasmid file: [[Media:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb|pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb]] (GenBank format)
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| |}
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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.
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