Difference between revisions of "Part:BBa K1725041"
 
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<partinfo>BBa_K1725041 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1725041 SequenceAndFeatures</partinfo>
  
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K1725000 (PhlF repressible promoter) driven expression is repressed by K1725040 as shown in the figure below. K1725042 is K1725040 driven by the <i>lacI</i> regulated promoter K1725080. Our control was K1725083 (the Tet repressor C0040 also driven by K1725080) and K1725082 (the TetR repressible promoter R0040 driving expression of I13500).
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PhlF gave 83-fold repression of GFP expression from PphlF (<bbpart>BBa_K1725000</bbpart>), whereas the control, TetR (<bbpart>BBa_C0040</bbpart>), gave only 33-fold repression of pL-tet (<bbpart>BBa_R0040</bbpart>). (figure 1) A lower expression level of PhlF completely represses GFP expression from PphlF, compared to the higher expression level of TetR required for pL-tet. (figure 2)
  
  
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https://static.igem.org/mediawiki/2015/9/95/Glasgow_2015_Repression_Fold_Graph.png
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https://static.igem.org/mediawiki/parts/7/78/Glasgow_2015_Fold_Repression_Graph.png
  
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<b>Represser constructs with pSB1C3 backbone; promoter driving GFP constructs with pSB3K3 backbone. Repressor protein expression induced with 100μM IPTG. Replicates of constructs and controls of three dilutions from one colony, under the same conditions. Mean and standard deviation of replicates were calculated to give value and error bars.</b>
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<b>Figure 1 Fold Repression. BioBricks: K1725082 (pL-tet driving GFP expression), K1725031 (pL-lac driving TetR expression), K1725001 (PphlF driving GFP expression), and K1725042 (pL-lac driving PhlF expression). Repressor protein expression induced with 100μM IPTG. Mean and standard deviation of replicates were calculated to give value and error bars, and normalised against a negative control.</b>
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https://static.igem.org/mediawiki/parts/6/6e/Glasgow_2015_Varied_IPTG_Graph.png
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<b>Figure 2 Varied Repression Levels. BioBricks: K1725082 (pL-tet driving GFP expression), K1725031 (pL-lac driving TetR expression), K1725001 (PphlF driving GFP expression), and K1725042 (pL-lac driving PhlF expression). Repressor protein expression induced with 100μM, 30 μM, 10 μM, 3 μM, and 0 μM IPTG. Mean and standard deviation of replicates were calculated to give value and error bars, and normalised against a negative control.</b>
  
 
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Latest revision as of 19:23, 20 September 2015

RBS + PhlF repressor + terminator

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

PhlF gave 83-fold repression of GFP expression from PphlF (BBa_K1725000), whereas the control, TetR (BBa_C0040), gave only 33-fold repression of pL-tet (BBa_R0040). (figure 1) A lower expression level of PhlF completely represses GFP expression from PphlF, compared to the higher expression level of TetR required for pL-tet. (figure 2)


Glasgow_2015_Fold_Repression_Graph.png

Figure 1 Fold Repression. BioBricks: K1725082 (pL-tet driving GFP expression), K1725031 (pL-lac driving TetR expression), K1725001 (PphlF driving GFP expression), and K1725042 (pL-lac driving PhlF expression). Repressor protein expression induced with 100μM IPTG. Mean and standard deviation of replicates were calculated to give value and error bars, and normalised against a negative control.

Glasgow_2015_Varied_IPTG_Graph.png

Figure 2 Varied Repression Levels. BioBricks: K1725082 (pL-tet driving GFP expression), K1725031 (pL-lac driving TetR expression), K1725001 (PphlF driving GFP expression), and K1725042 (pL-lac driving PhlF expression). Repressor protein expression induced with 100μM, 30 μM, 10 μM, 3 μM, and 0 μM IPTG. Mean and standard deviation of replicates were calculated to give value and error bars, and normalised against a negative control.