Regulatory

Part:BBa_K3198004

Designed by: Chunyang Song   Group: iGEM19_NUS_Singapore   (2019-08-23)
Revision as of 06:24, 4 October 2019 by Tiantian0412 (Talk | contribs)


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.


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