This part was designed to test the functionality of the constitutive promotor region.
<u><b>Characterization</b></u>
<u><b>Characterization</b></u>
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<p align="justify">We fused the constitutive promoter to a GFP reporter, and assayed the promoter’s activity after addition of different amounts of arabinose and IPTG. As such these results can serve as a control for the results obtained for the arabinose-inducible [https://parts.igem.org/wiki/index.php?title=Part:BBa_K584000 BBa_K584000] and IPTG-inducible [https://parts.igem.org/wiki/index.php?title=Part:BBa_K584002 BBa_K584002] bricks.
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We tested the activity both in a TOP10F’ (Figure 1 & 3) as well as a MG1655 (Figure 2 & 4) E.coli strain background. For more information on E.coli strain descriptions, we recommend the following website: [http://openwetware.org/wiki/E._coli_genotypes E.Coli_Genotypes].</p>
<p align="justify">Apart from a small growth defect and somewhat lower fluorescence at the 8 hour time point after the addition of 2% arabinose, the activity of the constitutive promoter in TOP10F’ cells is unaffected by arabinose addition, as seen in Figure1. For the MG1655 cells (Figure 2), however, we see clear effects of adding arabinose at concentrations of 0.2% or 2%, with promoter activities significantly lower than the control condition. This points to interference of high inducer levels with the activity of the promoter and the importance of metabolism of the inducer, as arabinose can be metabolized by MG1655 cells, but not by TOP10F’ cells.</p>
<p align="justify">For both TOP10F’ and MG1655 cells, the activity of the constitutive promoter is unaffected by the addition of IPTG (Figures 3 and 4).</p>
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<p align="justify">The <b>[http://2011.igem.org/Team:KULeuven K.U.Leuven 2011 iGEM Team]</b> has <b><u>extensively characterized</u></b> the <b><u>existing Biobrick part</u></b> [https://parts.igem.org/Part:BBa_J23119:Experience Constitutive promoter(BBa_J23119)].</p>
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[[Image:pConst arab MG1655.jpg|thumb|center|Figure4]]
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The detailed qualitative and quantitative <u><b>[https://parts.igem.org/Part:BBa_J23119:Experience review]</b></u> has been added into the experience page of the BBa_J23119 Constitutive promoter .
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Latest revision as of 10:00, 21 September 2011
Constitutive promoter + GFP generator
Characterization
The [http://2011.igem.org/Team:KULeuven K.U.Leuven 2011 iGEM Team] has extensively characterized the existing Biobrick partConstitutive promoter(BBa_J23119).
The detailed qualitative and quantitative review has been added into the experience page of the BBa_J23119 Constitutive promoter .
Sequence and Features
Assembly Compatibility:
10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7 Illegal NheI site found at 30
