Composite
Urea Senso

Part:BBa_K3710004

Designed by: Sandra Lizeth Martinez Diaz, Erik Hanko and Katherine Baker   Group: iGEM21_Manchester   (2021-10-01)
Revision as of 06:34, 1 October 2021 by Lizzzethmtz (Talk | contribs)

This part encodes the Escherichia coli codon optimized parts for a Urea biosensor genetic circuit comprising the urea-specific transcription factor UreR and the bidirectional promoter of the Proteus mirabilis urease operon pUreD.

Proteus mirabilis encodes the transcriptional regulator UreR of the Urease gene cluster belonging to the AraC/XylS family of transcriptional regulators. UreR upregulates the gene clusters’ subunit transcription and recruits the accessory proteins (UreDABCEFG), when accompanied by urea, to form an active urease [2].

Pathogenic bacteria produce virulence factors such as ureases to assist in their colonisation efforts. Consequently, these ureases hydrolyse urea into NH3 and Carbonic acid, which increase local pH and provide a processed nitrogen source for growth. These changes assist the bacteria but also harm the host, causing mineral deposition on the urethral lumen resulting in urinary stones [1].

pUreD is an inducible promoter that is activated by the UreR transcription factor in the presence of urea. pUreD is one of several promoters activated by the UreR transcriptional activator which share a consensus sequence of TA/GT/CA/TT/GC/TTA/TT/AATTG [3].

pUreD is a bidirectional promoter driving transcription of the UreR and UreD protein subunits of the urease operon of P. mirabilis. pUreD contains two ribosome binding sites and a UreR binding site about upstream of the forward transcriptional start site.


References: [1] Poore, C. and Mobley, H. (2003) "Differential regulation of the Proteus mirabilis urease gene cluster by UreR and H-NS", Microbiology, 149(12), pp. 3383-3394. doi: 10.1099/mic.0.26624-0. [2] Dattelbaum, J. et al. (2003) "UreR, the Transcriptional Activator of the Proteus mirabilis Urease Gene Cluster, Is Required for Urease Activity and Virulence in Experimental Urinary Tract Infections", Infection and Immunity, 71(2), pp. 1026-1030. doi: 10.1128/iai.71.2.1026-1030.2003.

[3]. Thomas V, Collins C. Identification of UreR binding sites in the Enterobacteriaceae plasmid-encoded and Proteus mirabilis urease gene operons. Molecular Microbiology. 1999;31(5):1417-1428.

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