Difference between revisions of "Part:BBa K2332002"

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Pblind is a blue light inducible promoter. It requires cells to also express EL222 ([https://parts.igem.org/Part:BBa_K2332004 BBa_K2332004]), a natural photosensitive DNA-binding protein that becomes active only upon blue-light exposure. This part consists of a fusion of the EL222 DNA binding region and the LuxI promoter. The lux box, a 20bp inverted repeat (LuxR and 3-oxo-C6-HSL complex binding region) from the luxI promoter, was replaced with the 18bp DNA binding region of EL222. In the dark, EL222 is inactive as its N-terminal LOV domain represses its DNA-binding C-terminal HTH domain. Upon blue light exposure (450nm), LOV-HTH interaction is released, allowing it to dimerize and bind its binding region, overlapping the -35 region of the luxI promoter. This ultimately results in the recruitment of RNAP and transcriptional activation.
 
Pblind is a blue light inducible promoter. It requires cells to also express EL222 ([https://parts.igem.org/Part:BBa_K2332004 BBa_K2332004]), a natural photosensitive DNA-binding protein that becomes active only upon blue-light exposure. This part consists of a fusion of the EL222 DNA binding region and the LuxI promoter. The lux box, a 20bp inverted repeat (LuxR and 3-oxo-C6-HSL complex binding region) from the luxI promoter, was replaced with the 18bp DNA binding region of EL222. In the dark, EL222 is inactive as its N-terminal LOV domain represses its DNA-binding C-terminal HTH domain. Upon blue light exposure (450nm), LOV-HTH interaction is released, allowing it to dimerize and bind its binding region, overlapping the -35 region of the luxI promoter. This ultimately results in the recruitment of RNAP and transcriptional activation.
  
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===Usage and Biology===
 
===Usage and Biology===
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[[File:Pblind-GFP.png|thumb|center|500px| Figure 1: Blue light inducible expression system. Under blue light, the EL222 DNA binding protein dimerises and bind its binding region within the designed Pblind promoter, overlapping the -35 region of the luxI promoter. This ultimately results in the recruitment of RNAP and transcriptional activation. Figure adapted from [https://www.ncbi.nlm.nih.gov/pubmed/27353329 Jayaraman P. et al. (2016)]]]
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Revision as of 23:11, 23 October 2017


Blue light inducible promoter (Pblind)

Pblind is a blue light inducible promoter. It requires cells to also express EL222 (BBa_K2332004), a natural photosensitive DNA-binding protein that becomes active only upon blue-light exposure. This part consists of a fusion of the EL222 DNA binding region and the LuxI promoter. The lux box, a 20bp inverted repeat (LuxR and 3-oxo-C6-HSL complex binding region) from the luxI promoter, was replaced with the 18bp DNA binding region of EL222. In the dark, EL222 is inactive as its N-terminal LOV domain represses its DNA-binding C-terminal HTH domain. Upon blue light exposure (450nm), LOV-HTH interaction is released, allowing it to dimerize and bind its binding region, overlapping the -35 region of the luxI promoter. This ultimately results in the recruitment of RNAP and transcriptional activation.


Usage and Biology

Figure 1: Blue light inducible expression system. Under blue light, the EL222 DNA binding protein dimerises and bind its binding region within the designed Pblind promoter, overlapping the -35 region of the luxI promoter. This ultimately results in the recruitment of RNAP and transcriptional activation. Figure adapted from Jayaraman P. et al. (2016)


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]