Difference between revisions of "Part:BBa K206001"

 
 
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__NOTOC__
 
__NOTOC__
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<partinfo>BBa_K206001 short</partinfo>
 
  
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Weaker version of wild type pBAD (<partinfo>I13453</partinfo>).
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{|
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|<div style="align: center; valign: center; font-family: Arial; font-size: 12pt">
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<!-- Add more about the biology of this part here
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{|
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|-
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|width="60px"|''Name'':
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|pBAD weak
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|-
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|width="60px"|''Input'':
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|width="100px"|[http://openwetware.org/wiki/Arabinose L-arabinose]
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|-
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|''Output'':
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| PoPS
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|}
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</div>
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<hr width="800px">
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<div class="noprint" style="padding: 10px; color: #ffffff; background-color: #C0C0C0; width: 800px; align: center">
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<center>
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[[Part:BBa_K206001 |Part Main Page]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[Part:BBa_K206001:Design |Part Design]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[Part:BBa_K206001:Characterization |Characterization]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[pBAD Promoter Family |Family]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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[[Part:BBa_K206001:Experience |Add Data]] &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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</center>
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</div>
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|}
 
===Usage and Biology===
 
===Usage and Biology===
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pBAD is an <i>E.coli</i> promoter that is induced by L-arabinose. In the absence of arabinose, the repressor protein AraC (<partinfo>I13458</partinfo>) binds to the AraI1 operator site of pBAD and the upstream operator site AraO2, blocking transcription [[Part:BBa_K206001#References|[1]]]. In the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription [[Part:BBa_K206001#References|[1]]].
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<br>
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<br>
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K206001 is a variant pBAD promoter with a modified AraI1 site that has been shown to be less responsive at low concentrations of arabinose and to exhibit a lower maximum expression than the wild type (<partinfo>I13453</partinfo>), as measured by [[Part:BBa_K206003|coupling to a fluorescent reporter]].
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<br>
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<br>
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''What you can do with it:''
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<br>
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At a given level of arabinose input, <partinfo>K206001</partinfo> will provide a lower level of PoPS output than its [[pBAD Promoter Family|family members]], allowing analog device responses. See [http://2009.igem.org/Team:British_Columbia our wiki] for a project that makes use of this property.
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<br>
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<br>
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''Compatibility'':
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<br>
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Chassis: Best used in the ''E. coli'' strain [http://cgsc.biology.yale.edu/Strain.php?ID=111773 BW27783], which has been modified to permit homogeneous pBAD promoter expression by substituting the chromosomal arabinose-dependent promoter of AraE (arabinose transporter protein) with a constitutive promoter [[Part:BBa_K206001#References|[2]]].
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<br>
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Backbone: Has been shown to work on plasmid <partinfo>pSB1C3</partinfo>.
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<br>
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Reporter: Has been shown to work with reporters <partinfo>I13507</partinfo> and <partinfo>I763020</partinfo>.
  
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<br>
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<span class='h3bb'>Sequence and Features</span>
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===<span class='h3bb'>Sequence and Features</span>===
 
<partinfo>BBa_K206001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K206001 SequenceAndFeatures</partinfo>
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<br>
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===References===
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[http://www.ncbi.nlm.nih.gov/pubmed/11102706 [1]] Schlief, R. (2000). Regulation of the L-arabinose operon of ''Escherichia coli''. Trends in Genetics. '''16'''(12):559-565.
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<br>
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[http://www.ncbi.nlm.nih.gov/pubmed/11739756 [2]] Khlebnikov A, Datsenko KA, Skaug T, Wanner BL, and Keasling JD. (2001). Homogeneous expression of the PBAD promoter in Escherichia coli by constitutive expression of the low-affinity high-capacity AraE transporter. Microbiology. '''147'''(12):3241-7.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 02:09, 22 October 2009


Name: pBAD weak
Input: [http://openwetware.org/wiki/Arabinose L-arabinose]
Output: PoPS

Part Main Page        Part Design        Characterization        Family        Add Data       

Usage and Biology

pBAD is an E.coli promoter that is induced by L-arabinose. In the absence of arabinose, the repressor protein AraC (BBa_I13458) binds to the AraI1 operator site of pBAD and the upstream operator site AraO2, blocking transcription [1]. In the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription [1].

K206001 is a variant pBAD promoter with a modified AraI1 site that has been shown to be less responsive at low concentrations of arabinose and to exhibit a lower maximum expression than the wild type (BBa_I13453), as measured by coupling to a fluorescent reporter.

What you can do with it:
At a given level of arabinose input, BBa_K206001 will provide a lower level of PoPS output than its family members, allowing analog device responses. See [http://2009.igem.org/Team:British_Columbia our wiki] for a project that makes use of this property.

Compatibility:
Chassis: Best used in the E. coli strain [http://cgsc.biology.yale.edu/Strain.php?ID=111773 BW27783], which has been modified to permit homogeneous pBAD promoter expression by substituting the chromosomal arabinose-dependent promoter of AraE (arabinose transporter protein) with a constitutive promoter [2].
Backbone: Has been shown to work on plasmid pSB1C3.
Reporter: Has been shown to work with reporters BBa_I13507 and BBa_I763020.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[http://www.ncbi.nlm.nih.gov/pubmed/11102706 [1]] Schlief, R. (2000). Regulation of the L-arabinose operon of Escherichia coli. Trends in Genetics. 16(12):559-565.
[http://www.ncbi.nlm.nih.gov/pubmed/11739756 [2]] Khlebnikov A, Datsenko KA, Skaug T, Wanner BL, and Keasling JD. (2001). Homogeneous expression of the PBAD promoter in Escherichia coli by constitutive expression of the low-affinity high-capacity AraE transporter. Microbiology. 147(12):3241-7.