Difference between revisions of "Part:BBa K3046014"

 
 
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<partinfo>BBa_K3046014 short</partinfo>
 
<partinfo>BBa_K3046014 short</partinfo>
  
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This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project
  
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===Usage and Biology===
 
===Usage and Biology===
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This is a promoter for <i>Aspergillus niger</i> of unknown strength that should be active in the stationary phase.
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===Characterization===
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<html>
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This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. [1] This version is a consensus sequence of an unknown gene's promoters in <i>Aspergillus</i>.
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<br>
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This promoter was characterised using an mCherry test device,<a href=”https://parts.igem.org/wiki/index.php?title=Part:BBa_K3046009” target=”_blank”>BBa_K3046009</a>, inserted into an AMA1-based test plasmid, <a href=”https://parts.igem.org/wiki/index.php?title=Part:BBa_K3046021” target=”_blank_”>BBa_K3046021</a>, and characterisation was done in a microbioreactor (BioLector, m2p-labs) for microtiter scale.
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<br><br>
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The predicted behavior of the consensus promoter, as described by the model, is summarized by the figure below. This promoter is expected to show a much lower expression, but maintain the same characteristics with regards to growth phase dependency.
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<br>
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<img src="https://static.igem.org/mediawiki/parts/b/bf/T--DTU-Denmark--unk_4_RNAseq.png" style="width: 80%; padding: 15px;" >
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<figure>
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<figcaption> Figure 1: The figure shows RNA-seq data for <i>Aspergillus niger</i> in both exponential and stationary phase with the glaA gene marked in red. The x-axis is the promoter activity in the stationary phase and the y-axis is the promoter activity in the exponential phase, both axes are depicted on a log scale. Here we see that the promoter should be moderately active in the stationary phase.
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</figcaption>
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</figure>
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<br>
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For the microtiter scale, the promoter was evaluated in a biolector, producing the following results with regards to growth, fluorescence and dynamic promoter activity.<br>
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<img src="https://static.igem.org/mediawiki/parts/f/f2/T--DTU-Denmark--unk_4.svg" style="width: 60%; padding: 15px;" >
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<figure>
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<figcaption> Figure 2: When analyzed in the microtiter scale on a biolector, the red fluorescence and biomass has been measured and the Dynamic Promoter Activity has been calculated. On the graph Dynamic Promoter Activity (DPA) is green, the biomass is measured in Light Scattering Units (LSU) is in blue, and red fluorescence (RFP) is shown in red. The graphs have been normalized for LSU.
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Here we see no appreciable promoter activity
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</figcaption>
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</figure>
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</html>
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Latest revision as of 21:11, 21 October 2019


PLEAPunk_4

This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project

Usage and Biology

This is a promoter for Aspergillus niger of unknown strength that should be active in the stationary phase.

Characterization

This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. [1] This version is a consensus sequence of an unknown gene's promoters in Aspergillus.
This promoter was characterised using an mCherry test device,BBa_K3046009, inserted into an AMA1-based test plasmid, BBa_K3046021, and characterisation was done in a microbioreactor (BioLector, m2p-labs) for microtiter scale.

The predicted behavior of the consensus promoter, as described by the model, is summarized by the figure below. This promoter is expected to show a much lower expression, but maintain the same characteristics with regards to growth phase dependency.

Figure 1: The figure shows RNA-seq data for Aspergillus niger in both exponential and stationary phase with the glaA gene marked in red. The x-axis is the promoter activity in the stationary phase and the y-axis is the promoter activity in the exponential phase, both axes are depicted on a log scale. Here we see that the promoter should be moderately active in the stationary phase.

For the microtiter scale, the promoter was evaluated in a biolector, producing the following results with regards to growth, fluorescence and dynamic promoter activity.
Figure 2: When analyzed in the microtiter scale on a biolector, the red fluorescence and biomass has been measured and the Dynamic Promoter Activity has been calculated. On the graph Dynamic Promoter Activity (DPA) is green, the biomass is measured in Light Scattering Units (LSU) is in blue, and red fluorescence (RFP) is shown in red. The graphs have been normalized for LSU. Here we see no appreciable promoter activity


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 42
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 402
  • 1000
    COMPATIBLE WITH RFC[1000]