BBa_K352004

BBa_K352004 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K352004
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Bilgi Gungor
Date created: 2010-10-01 11:00:00
Date modified: 2015-05-08 01:12:11

G2C Mutated pCooF from Rhodospirillum rubrum



Types
DnaRegion

Roles
promoter

Regulatory

Sequences BBa_K352004_sequence (Version 1)

Description

This part is the mutated version of pCooF promoter. The mutation involves conversion guanine to cytosine at second base. The binding strength of pCooF to CooA is expected to diminish.

CooA is a CO-sensing protein that activates the transcription of genes encoding the CO-oxidation (coo) regulon, whose polypeptide products are required for utilizing CO as an energy source in Rhodospirillum rubrum. CooA binds to a position overlapping the -35 element of the P(cooF) promoter, similar to the arrangement of class II CRP (cAMP receptor protein)- and FNR (fumarate and nitrate reductase activator protein)-dependent promoters when expressed in Escherichia coli.
The CO-dependent anaerobic growth of Rhodospirillum rubrum relies on a CO oxidation system encoded by two CO regulated transcriptional units, cooMKLXUH and cooFSCTJ. The key products of the coo regulon are an O2- sensitive CO dehydrogenase (CooS), a CooS-associated Fe-S protein (CooF), and a CO-tolerant hydrogenase (CooH), and the expression of the genes depends upon the activity of the CooA protein, which senses CO under anaerobic conditions.

CO-independent basal level transcription of PcooF is not detectable due to the absence of active CooA.

Notes

The sequence information was acquired from NCBI and physical DNA was synthesized from GENEART. Classical cloning strategies(restriction digestion) were used to produce biobricks

Source

de Vooght KM, van Wijk R, van Solinge WW. Management of gene promoter mutations in molecular diagnostics. Clin. Chem.55(4),698???708 (2009)

Sequence Annotation Location Component / Role(s)
CooA Binding Site
Transcription Start Site
CooA Binding Site
-10 Region
Protected by CooA+RNAP
A+T Rich Sequence
Protected by CooA
41,44
92,93
52,58
79,86
23,91
8,33
36,61
feature/binding non_covalent_binding_site
feature/start start_codon
feature/binding non_covalent_binding_site
feature/misc sequence_feature
conserved_region feature/conserved
sequence_feature feature/misc
conserved_region feature/conserved
igem#experience
None
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K352004/1