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.
|- valign=top
| P70
| Lambda phage promoter OR2-OR1-Pr specific to E. coli σ70
| GenBank: J02459.1
|- valign=top
|- valign=top
| 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.

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.

  • Raw data file: plasmid_deGFP-070812.xls (Excel spreadsheet)
  • Information about protocol on "Outline" sheet; plotted data is on the sheet "plotted-data"


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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