This page contains a description of the DNA parts used for the biomolecular breadboards. These parts will be submitted to Addgene (eventually).
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.
|| Lambda phage promoter OR2-OR1-Pr specific to E. coli σ70
|| GenBank: J02459.1
|| The untranslated region containing the T7 g10 leader sequence for highly efficient translation initiation
|| GenBank: M35614.1
|| 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.
|| 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.
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.