Difference between revisions of "Part:BBa K1065001"

 
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<partinfo>BBa_K1065001 short</partinfo>
 
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This is a composite part composed by an AraC-pBAD promoter (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K731201">BBa_K731201</a>) + 2-oxoglutarate oxygenase/decarboxylase (<a href="https://parts.igem.org/Part:BBa_K1065000">BBa_K1065000</a>). This construct allowed us to characterize the Ethylene Forming Enzyme, inducing its expression using Arabinose at a concentration of 5 mM.
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===EFE characterization===
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In order to obtain a good characterization of the part, a set of experiments were done.
  
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===Usage and Biology===
 
  
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<h3>Effect of EFE on cell growth</h3>
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<center><img src="https://static.igem.org/mediawiki/2013/6/6f/Tn-20130627-Efe_Toxicity_test-PLOT.png"></center><br/>
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<p style="width:600px; margin-left:150px; margin-bottom:60px; text-align:justify"><b>Figure 1</b> Effect of EFE on cell growth. Cell density was measured at different time points to determine the effect of EFE expression. Cells were grown at 37 °C in LB until it was reached an OD of 0.6. The cells were then slitted in four samples of equal volume. Two samples were then induced with 5 mM Arabinose. Induced samples show a slowed growth rate,
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as espected (5mM arabinose is a strong induction that causes stress on cells). However, cell growth is not completely inhibited so EFE is not highly toxic</p>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 08:23, 8 August 2013

AraC-pBAD + 2-oxoglutarate oxygenase/decarboxylase (EFE) + terminators

This is a composite part composed by an AraC-pBAD promoter (BBa_K731201) + 2-oxoglutarate oxygenase/decarboxylase (BBa_K1065000). This construct allowed us to characterize the Ethylene Forming Enzyme, inducing its expression using Arabinose at a concentration of 5 mM. ===EFE characterization=== In order to obtain a good characterization of the part, a set of experiments were done.

Effect of EFE on cell growth



Figure 1 Effect of EFE on cell growth. Cell density was measured at different time points to determine the effect of EFE expression. Cells were grown at 37 °C in LB until it was reached an OD of 0.6. The cells were then slitted in four samples of equal volume. Two samples were then induced with 5 mM Arabinose. Induced samples show a slowed growth rate, as espected (5mM arabinose is a strong induction that causes stress on cells). However, cell growth is not completely inhibited so EFE is not highly toxic

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1530
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1228
    Illegal AgeI site found at 979
    Illegal AgeI site found at 2281
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961