Biomolecular Breadboards:DNA parts: Difference between revisions
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{{Template:Biomolecular Breadboards}} | {{Template:Biomolecular Breadboards}} | ||
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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. | |||
{| | |||
|- valign=top | |||
| [[Image:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.png]] | |||
| | |||
* Control sequence: [[Media:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb|pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb]] (GenBank file) | |||
* Preliminary data: [[Media:plasmid_deGFP-070812.xls|plasmid_deGFP-070812.xls]] (Excel spreadsheet) | |||
|} | |||
=== Plasmid description === | |||
{| border=1 width=100% cellpadding=4 | |||
|- valign=top | |||
| P70 | |||
| Lambda phage promoter OR2-OR1-Pr specific to E. coli σ70 | |||
| GenBank: J02459.1 | |||
|- valign=top | |||
| UTR1 | |||
| The untranslated region containing the T7 g10 leader sequence for highly efficient translation initiation | |||
| GenBank: M35614.1 | |||
|- valign=top | |||
| 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. | |||
|- valign=top | |||
| 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. | |||
{| | |||
|- | |||
| | |||
[[Image:plasmid_sat.png|400px]] | |||
| valign=top | | |||
* Raw data file: [[Media:plasmid_deGFP-070812.xls|plasmid_deGFP-070812.xls]] (Excel spreadsheet) | |||
* Information about protocol on "Outline" sheet; plotted data is on the sheet "plotted-data" | |||
<hr> | |||
* Control plasmid file: [[Media:pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb|pBEST_OR2_OR1_Pr_UTR-deGFP-T500.gb]] (GenBank format) | |||
|} | |||
'''Figure 1. eGFP expression as a function of plasmid DNA template.''' Plasmid DNA pBEST-OR2-OR1-Pr-UTR1-deGFP-T500 is varied by concentration. |
Revision as of 21:08, 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.