Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Amanda Urke
Date created: 2016-12-11 12:00:00
Date modified: 2016-12-11 05:29:45
Glucose Binding Protein of Thermus thermophilus
| Types | DnaRegion |
| Roles | polypeptide_domain Protein_Domain |
| Sequences | BBa_M50037_sequence (Version 1) |
Description
This bacterial glucose binding protein (GBP) is derived from the genome sequence of Thermus thermophilus that was published in 2004 by Henne et al. In 2006, Cuneo et al. analyzed the sequence of this particular GBP, determined its X-ray crystal structure and characterized the stability of glucose-binding. The GBP is composed of two domains separated by a hinge, maintaining an open conformation when unbound to glucose and a closed conformation when bound. When flanked with an RFP fluorophore on the N-terminus and a GFP fluorophore on the C-terminus, these conformation changes facilitated FRET-based analysis of glucose binding. In addition, this GBP was selected for its reasonably short sequence length (1242bp) and stable glucose binding ability (Kd value of 0.08(+/- 0.03) mM). According to Cuneo et al., this T. thermophilus GBP shares more structural homology with maltose binding proteins than glucose binding proteins. However, we were able to obtain FRET when glucose was added, suggesting that glucose is a ligand for this protein.Notes
This GBP sequence was optimized for use in E. coli. Also, start and stop codons were removed to facilitate fusion to the other components of our construct.Source
This GBP comes from the genomic sequence of Thermus thermophilus. The complete bacterial genome was published by Henne et al. in Nature Biotechnology in 2004 (https://www.ncbi.nlm.nih.gov/pubmed/15064768), and the particular GBP was characterized in a 2006 study by Cuneo et al. in the Journal of Molecular Biology (https://www.ncbi.nlm.nih.gov/pubmed/16904687).| Sequence Annotation | Location | Component / Role(s) |
| Deletion of start codon to facilitate fusion to linker and fluorophore Deletion of stop codon to facilitate fusion to linker and fluorophore | 1,1 1239,1239 | start_codon feature/start feature/stop stop_codon |