Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jonathan Deaton
Date created: 2014-10-22 11:00:00
Date modified: 2015-05-08 01:14:03
HeLa Cell Glutathione Sensor
| Types | DnaRegion |
| Roles | Project engineered_region |
| Sequences | BBa_M36091_sequence (Version 1) |
Description
This part is designed to sense the presence of glutathione. It is designed to be used in HeLa cells. The first two elements (at the 5??? end) of this part consist of a constitutive mammalian Promoter (PGK) and a kozak sequence, which are designed to together constitutively express PAX-8. PAX-8 is a cis-element transcription factor that activates the thyroglobulin promoter (tgbp). PAX-8 activity is upregulated in the presence of glutathione. Directly downstream of the PAX-8 gene is a Stop Codon, a Poly-A Tail Signal Sequence, and the Thyroglobulin Promoter (TGp). The design of this part was derived primarily from Fig. 1-15 in "Ontogeny, Anatomy, Metabolism and Physiology of the Thyroid" (Dumont et al. 2011) and "Novel roles for glutathione in gene expression, cell death, and membrane transport of organic solutes" (Hammond et al. 2001), as well as sequences sources online.Notes
The main restriction in this construct was keeping DNA length under 2500 base pairs. This required removal of a Thyroglobulin (TG) Enhancer sequence 1902 to 1744 base pairs upstream of TG transcription Initiation. This size limit also required us to eliminate the Transcription Termination Factor-1 (TTF-1), another transcription factor important in the activation of the TG promoter.Source
http://www.journal-of-hepatology.eu/article/S0168-8278(01)00037-X/pdfhttp://www.thyroidmanager.org/chapter/ontogeny-anatomy-metabolism-and-physiology-of-the-thyroid/