Difference between revisions of "Part:BBa K3071013"
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<center><u>Figure 3 illustration of our synthetic biological system upon DSF activation</u></center> | <center><u>Figure 3 illustration of our synthetic biological system upon DSF activation</u></center> | ||
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− | The UAS sites of this promoter are replaced by CBS I( | + | The UAS sites of this promoter are replaced by CBS I((<html><a href="https://parts.igem.org/Part:BBa_K3071011">BBa_K3071011</a></html>) and CBSII (<html><a href="https://parts.igem.org/Part:BBa_K3071012">BBa_K3071012</a></html>) in our own synthetic system to construct CBS I & II-regulated pspA promoter (<html><a href="https://parts.igem.org/Part:BBa_K3071014">BBa_K3071014</a></html>). |
<center>[[file:T--Hong_Kong-CUHK--DSF_assay.png]]</center> | <center>[[file:T--Hong_Kong-CUHK--DSF_assay.png]]</center> |
Revision as of 07:09, 21 October 2019
PspA promoter
Description
This is an regulatory element that originated from Escherichia coli K12 strain. This basic part is used to construct our composite part (BBa_K3071014). Our rt-qPCR data show that this promoter is able to initiate the transcription in form of synthetic construct.
Biology
pspA promoter is a sigma-54 (σ-54) regulated activator dependent promoter. In the orginal pspA promoter upstream region, it contains σ-54 consensus sequence 5' of the start contains a GG doublet as -24 and a consensus GG doublet at -12, the high-affinity IHF site (-25 to -60), as well as the UAS sites (UAS I: -89 to -107; UAS II:-111 to -129).
Usage
sigm54-RNA holoenzyme (σ-54 RNAP) forms an inactive transcriptional initiation complex on this promoter, which can be activated in E. coli by the bacterial enhancer-binding protein PspF (BBa_K3071006). PspF functions by binding to the upstream activation sequences (UAS) near the promoter and contacting the promoter-bound σ-54 RNAP via DNA looping stabilized by the binding of integration host factor (IHF). Previous research has demonstrated the property of pspF-dependent and enhancer-specific transcription activation of pspA promoter.
Characterization
The UAS sites of this promoter are replaced by CBS I((BBa_K3071011) and CBSII (BBa_K3071012) in our own synthetic system to construct CBS I & II-regulated pspA promoter (BBa_K3071014).
The rt-qPCR data upon activation by DSF indicate a significant change in gene expression, suggest the related composite part is functional.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 12
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]