Difference between revisions of "Part:BBa K3198004"

 
 
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<partinfo>BBa_K3198004 short</partinfo>
 
<partinfo>BBa_K3198004 short</partinfo>
  
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Team iGEM 18 NUS Singapore-A submitted BBa_K2819103 which is a Blue light-repressible system with RFP reporter attached with YbaQ degradation tag.
 
  
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This part contains a promoter that can be repressed by blue light (450nm). This promoter makes use of a blue light dependent DNA-binding protein, EL222. Irradiation by blue light of wavelength 450nm exposes the hitherto sequestered HTH, facilitating dimerization of EL222 and subsequent DNA binding. The repression is achieved by placing the DNA binding site of EL222 between the -35 and -10 hexamers of the consensus promoter in E. coli, creating the blue light repressible promoter PBLrep. As a result, EL222 acts as a repressor, blocking the binding of RNA polymerase and repress gene expression in the presence of blue light. In the dark, RNA polymerase can now bind, and gene expression takes place.
 
  
  
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3198004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3198004 SequenceAndFeatures</partinfo>
  
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===Description===
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This part contains a promoter that can be repressed by blue light (450nm). This promoter makes use of a blue light dependent DNA-binding protein, EL222. Irradiation by blue light of wavelength 450nm exposes the hitherto sequestered HTH, facilitating dimerization of EL222 and subsequent DNA binding. The repression is achieved by placing the DNA binding site of EL222 between the -35 and -10 hexamers of the consensus promoter in <i>Escherichia coli</i>, creating the blue light repressible promoter PBLrep. As a result, EL222 acts as a repressor, blocking the binding of RNA polymerase and repress gene expression in the presence of blue light. In the dark, RNA polymerase can now bind, and gene expression takes place.
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<br><br>This year, we constructed two different new plasmids. One of them had an increased spacer DNA sequence with 30 base pairs, while the other one had the spacer sequence removed completely. Our aim was to investigate how the length of this spacer DNA would affect the performance of the blue-light repressible system.
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===Usage&Biology===
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This part contains a promoter that can be repressed by blue light (450nm). This promoter makes use of a blue light dependent DNA-binding protein, EL222. Irradiation by blue light of wavelength 450nm exposes the hitherto sequestered HTH, facilitating dimerization of EL222 and subsequent DNA binding. The repression is achieved by placing the DNA binding site of EL222 between the -35 and -10 hexamers of the consensus promoter in <i>Escherichia coli</i>, creating the blue light repressible promoter PBLrep. As a result, EL222 acts as a repressor, blocking the binding of RNA polymerase and repress gene expression in the presence of blue light. In the dark, RNA polymerase can now bind, and gene expression takes place.
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===Improvement over existing part [https://parts.igem.org/Part:BBa_K2819103 BBa_K2819103] by iGEM18 NUS Singapore-A===
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===Characterization===
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Please go to https://parts.igem.org/wiki/index.php?title=Part:BBa_K3198005 for further details.
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3198004 parameters</partinfo>
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<partinfo>BBa_K3198001 parameters</partinfo>
 
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Latest revision as of 09:58, 17 October 2019


Blue light repressible promoter with spacer addition



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]

Description

This part contains a promoter that can be repressed by blue light (450nm). This promoter makes use of a blue light dependent DNA-binding protein, EL222. Irradiation by blue light of wavelength 450nm exposes the hitherto sequestered HTH, facilitating dimerization of EL222 and subsequent DNA binding. The repression is achieved by placing the DNA binding site of EL222 between the -35 and -10 hexamers of the consensus promoter in Escherichia coli, creating the blue light repressible promoter PBLrep. As a result, EL222 acts as a repressor, blocking the binding of RNA polymerase and repress gene expression in the presence of blue light. In the dark, RNA polymerase can now bind, and gene expression takes place.

This year, we constructed two different new plasmids. One of them had an increased spacer DNA sequence with 30 base pairs, while the other one had the spacer sequence removed completely. Our aim was to investigate how the length of this spacer DNA would affect the performance of the blue-light repressible system.

Usage&Biology

This part contains a promoter that can be repressed by blue light (450nm). This promoter makes use of a blue light dependent DNA-binding protein, EL222. Irradiation by blue light of wavelength 450nm exposes the hitherto sequestered HTH, facilitating dimerization of EL222 and subsequent DNA binding. The repression is achieved by placing the DNA binding site of EL222 between the -35 and -10 hexamers of the consensus promoter in Escherichia coli, creating the blue light repressible promoter PBLrep. As a result, EL222 acts as a repressor, blocking the binding of RNA polymerase and repress gene expression in the presence of blue light. In the dark, RNA polymerase can now bind, and gene expression takes place.

Improvement over existing part BBa_K2819103 by iGEM18 NUS Singapore-A

Characterization

Please go to https://parts.igem.org/wiki/index.php?title=Part:BBa_K3198005 for further details.