Difference between revisions of "Part:BBa K3071013"
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<partinfo>BBa_K3071013 short</partinfo>
 
<partinfo>BBa_K3071013 short</partinfo>
 
===Description===
 
===Description===
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This is an regulatory element that originated from <i>Escherichia coli</i> K12 strain.
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This is an regulatory element that originated from <i>Escherichia coli</i> K12 strain. This basic part is used to construct our composite part ([https://parts.igem.org/Part:BBa_K3071014 BBa_K3071014]). Our rt-qPCR data show that this promotor is able to initiate the transcription in form of synthetic construct.
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===Biology===
 
===Biology===
 
<center>[[File:T--Hong_Kong-CUHK--pspA_promotor_structure.png]]</center>
 
<center>[[File:T--Hong_Kong-CUHK--pspA_promotor_structure.png]]</center>

Revision as of 10:04, 19 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 promotor is able to initiate the transcription in form of synthetic construct.

Biology

T--Hong Kong-CUHK--pspA promotor structure.png
Figure 1 Basic properties of pspA promotor

pspA promotor is a sigma-54 (σ-54) regulated activator dependent promotor. In the 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

T--Hong Kong-CUHK--sigma 54 activation.png
Figure 2 The transcription initiation mechanism of the sigma-54 promotors [http://www.cchem.berkeley.edu/wemmer/research/sigma54.html]


sigm54-RNA holoenzyme (σ-54 RNAP) forms an inactive transcriptional initiation complex on this promotor, 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 promotor-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 promotor.

Characterization

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 12
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]