Difference between revisions of "Part:BBa K203119"

 
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<partinfo>BBa_K203119 short</partinfo>
 
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A synthetic promoter upregulated by NF-&kappa;B activation. Manufactured by [http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters RA-PCR].
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===Usage and Biology===
 
===Usage and Biology===
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NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) is a transcription factor (TF) which regulates many different target genes resulting in the expression of various proteins. In most cell types (with the exception of B cells and Dentritic cells) NF-κB is bound to the Inhibitor of κB (IκB), which withhold NF-κB from entering the nucleus. When the cell becomes activated by an extra cellular stimluli, IκB is degraded and NF-κB can enter the nucleus. Within the nucleus NF-κB is able to enhance transcription of genes which are involved in immune response, cell proliferation or cell survival, depending on cell type and extra cellular stimuli [11]. In many cells NF-κB regulates anti-apoptotic proteins (e.g. TRAF1/2) and therby preventing cell death. Therefore mutations of NF-κB resulting in a constitutively active form are often associated with unregulated cell proliferation and cancer [12]. In macrophages the NF-κB signalling pathway could be activated by binding of bacterial lipopolysacchride (LPS). There NF-κB activation leads to secretion of cytokines which influence other lymphocytes.
  
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K203119 parameters</partinfo>
 
<partinfo>BBa_K203119 parameters</partinfo>
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NF-&kappa;B was induced with 2,5µM TNF-&alpha; in [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#U2-OS] cells. Expression level was determined in [http://2009.igem.org/Team:Heidelberg/Measurement REU] by flow cytometry (3 experiments on different days, 3 replicates / wells per experiment) 10 hours after induction.
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|[[Image:HD09_NFkB_31.png|thumb|none|350px|'''BBa_K203119 is a promoter induced approximately by 100% by TNF-&alpha;'''.  [http://2009.igem.org/Team:Heidelberg/Measurement REU] was determined by flow cytometry, 3 experiments à three replicates]]
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|[[Image:HD09_microscopy_NIIL10timepoints.PNG|thumb|none|350px|'''Timecourse of induction'''. Measured by image analysis. See [http://2009.igem.org/Team:Heidelberg/Project_Measurement#Flow_Cytometry/Fluorescence_Microscopy discussion] for why the results obtained by the two methods deviate.]]
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Revision as of 12:28, 16 October 2009

NfKB Responsive promoter

A synthetic promoter upregulated by NF-κB activation. Manufactured by [http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters RA-PCR].


Usage and Biology

NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) is a transcription factor (TF) which regulates many different target genes resulting in the expression of various proteins. In most cell types (with the exception of B cells and Dentritic cells) NF-κB is bound to the Inhibitor of κB (IκB), which withhold NF-κB from entering the nucleus. When the cell becomes activated by an extra cellular stimluli, IκB is degraded and NF-κB can enter the nucleus. Within the nucleus NF-κB is able to enhance transcription of genes which are involved in immune response, cell proliferation or cell survival, depending on cell type and extra cellular stimuli [11]. In many cells NF-κB regulates anti-apoptotic proteins (e.g. TRAF1/2) and therby preventing cell death. Therefore mutations of NF-κB resulting in a constitutively active form are often associated with unregulated cell proliferation and cancer [12]. In macrophages the NF-κB signalling pathway could be activated by binding of bacterial lipopolysacchride (LPS). There NF-κB activation leads to secretion of cytokines which influence other lymphocytes.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters

NF-κB was induced with 2,5µM TNF-α in [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#U2-OS] cells. Expression level was determined in [http://2009.igem.org/Team:Heidelberg/Measurement REU] by flow cytometry (3 experiments on different days, 3 replicates / wells per experiment) 10 hours after induction.

BBa_K203119 is a promoter induced approximately by 100% by TNF-α. [http://2009.igem.org/Team:Heidelberg/Measurement REU] was determined by flow cytometry, 3 experiments à three replicates
Timecourse of induction. Measured by image analysis. See [http://2009.igem.org/Team:Heidelberg/Project_Measurement#Flow_Cytometry/Fluorescence_Microscopy discussion] for why the results obtained by the two methods deviate.