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Team:UTK-Knoxville/HemAT
From 2013.igem.org
HemAT
HemAT
Oxygen gas, sensor FixL or HemAT (also need HemC). list of Direct Oxygen sensors . FixL, was fused with 5 different kinases : “it was likely that the autophosphorylation activity of the histidine kinase domain could not be regulated by the O2 association/dissociation in the sensor domain in the chimeric proteins” Intracellular & redox sensor. O2 sensor, would be interesting. & may also be able to sense CO in the absence of oxygen It seems like FixL is an O2 sensor.
Sensing domain is from 1-186, its cytoplasmic, so it might not work on EnvZ. It also does not HAMP domain so we do not think it would work on a transmembrane kinase domain.
Chimeric sensory kinases containing O2 sensor domain of FixL and histidine kinase domain from thermophile.
Oxygen‐Sensing Histidine‐Protein Kinases: Assays of Ligand Binding and Turnover of Response‐Regulator Substrates.
Heme-based oxygen sensor protein FixL: its structure and function.
Amino Acid Sequence:
MLFKKDRKQETAYFSDSNGQQKNRIQLTNKHADVKKQLKMVRLGDAELYVLEQLQPLIQENIVNIVDAFYKNLDHESSL
MDIINDHSSVDRLKQTLKRHIQEMFAGVIDDEFIEKRNRIASIHLRIGLLPKWYMGAFQELLLSMIDIYEASITNQQELLKA
IKATTKILNLEQQLVLE
KanR
KanR provides kanamycin resistance to colonies with successful transformations.
pBR322
The pBR322 ori is a low copy origin of replication derived from
NtrB
Details for NtrB coming soon.Details for NtrB coming soon.Details for NtrB coming soon.
NtrC
Details for NtrC coming soon.Details for NtrC coming soon.Details for NtrC coming soon.
T7 Promoter
The T7 promoter is constitutively expressed by the phage T7 RNA polymerase. The polymerase is orthogonal to the wildtype E. coli promoters.
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Gel samples
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Flow sample
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