Difference between revisions of "Part:BBa K1065001"
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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. | + | 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. </html> |
===EFE characterization=== | ===EFE characterization=== | ||
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1530
Illegal BamHI site found at 1144 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1228
Illegal AgeI site found at 979
Illegal AgeI site found at 2281 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961