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=== | ||
+ | [[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
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]