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| | This is a DNA sequence coding lanthanide binding tag described in literature. It's literatural dissociation constants are as follows:</br> | | This is a DNA sequence coding lanthanide binding tag described in literature. It's literatural dissociation constants are as follows:</br> |
| | K<sub>Tb<sup>3+</sup></sub>=18 nM</br> | | K<sub>Tb<sup>3+</sup></sub>=18 nM</br> |
| - | This is the lowest known value of dissociation constant, thus making the binding strenght highest amongst known LBTs.</br> | + | This is the lowest known value of dissociation constant for a Tb<sup>3+</sup>, thus making the binding strenght highest amongst known LBTs.</br> |
| | <h3>BBa_K1459013 - wSE3 lanthanide binding tag</h3></br> | | <h3>BBa_K1459013 - wSE3 lanthanide binding tag</h3></br> |
| | This is sequence of DNA coding wSE3 lanthanide binding tag. It's dissociation constants are as follows:</br> | | This is sequence of DNA coding wSE3 lanthanide binding tag. It's dissociation constants are as follows:</br> |
| - | K(Tb)=2000 nM</br> | + | K<sub>Tb<sup>3+</sup></sub>=2000 nM</br> |
| | <h3>BBa_K1459014 - Lanthanide Binding Tag</h3></br> | | <h3>BBa_K1459014 - Lanthanide Binding Tag</h3></br> |
| | This is DNA sequence coding a lanthanide binding tag. This one is one of the best described LBTs in literature, with dissociation constants following:</br> | | This is DNA sequence coding a lanthanide binding tag. This one is one of the best described LBTs in literature, with dissociation constants following:</br> |
| - | K(La)= 3500 nM</br> | + | K<sub>La<sup>3+</sup></sub>= 3500 nM</br> |
| - | K(Ce)= 950 nM</br> | + | K<sub>Ce<sup>3+</sup></sub>= 950 nM</br> |
| - | K(Nd)= 270 nM</br> | + | K<sub>Nd<sup>3+</sup></sub>= 270 nM</br> |
| - | K(Eu)= 62 nM</br> | + | K<sub>Eu<sup>3+</sup></sub>= 62 nM</br> |
| - | K(Gd)= 84 nM</br> | + | K<sub>Gd<sup>3+</sup></sub>= 84 nM</br> |
| - | K(Tb)= 57 nM</br> | + | K<sub>Tb<sup>3+</sup></sub>= 57 nM</br> |
| - | K(Dy)= 71 nM</br> | + | K<sub>Dy<sup>3+</sup></sub>= 71 nM</br> |
| - | K(Er)= 78 nM</br> | + | K<sub>Er<sup>3+</sup></sub>= 78 nM</br> |
| - | K(Yb)= 100 nM</br> | + | K<sub>Yb<sup>3+</sup></sub>= 100 nM</br> |
| - | K(Lu)= 128 nM</br> | + | K<sub>Lu<sup>3+</sup></sub>= 128 nM</br> |
| | <h3>BBa_K1459015 - 1L2Y short peptide</h3></br> | | <h3>BBa_K1459015 - 1L2Y short peptide</h3></br> |
| | This is DNA sequence coding short peptide (PDB 1L2Y) is highly structured in water and could provide a structural foundation for small binding tags, such as we were planning to use it.<br><br> | | This is DNA sequence coding short peptide (PDB 1L2Y) is highly structured in water and could provide a structural foundation for small binding tags, such as we were planning to use it.<br><br> |
Achivements
Parts
BBa_K1459001 - PmrA
Transcriptional factor PmrA takes part in iron (III) binding in
Salmonella enterica. Upon phosphorylation by PmrB it binds to pmrC promoter and induces expression of chemotaxis protein CheZ.
SENT TO REGISTRY
BBa_K1459002 - C-term of PmrB from Salmonella enterica
PmrB is a transmembrane kinase. After binding iron (III) ion by binding peptide on extracellular loop, it's intracellular domain gains kinase activity and phosphorylates PmrA (BBa_K1459000).
PmrB C-term is a part of two-component system. When fused with some binding tag, PmrB(N-term), PmrA and pmrC-reporter, it is a viable detecting system.
SENT TO REGISTRY
BBa_K1459003 - PmrA-PmrB(LBT) two-component system
PmrA-PmrB two-component system is native to
Salmonella enterica and in it's native state it is responsible for chemotaxis. PmrB is a transmembrane protein with iron binding peptide on it's extracellular loop. When PmrB binds iron (III) iron, it's intracellular domain gains kinase activity and phosphorylates PmrA, which subsequently binds to pmrC promoter and induces expression of chemotaxis CheZ protein. In this part iron binding tag on the extracellular loop was exchanged with a lanthanide binding tag (LBT), to allow PmrA-PmrB two-component system to respond to lanthanide ions.
BBa_K1459004 - PmrA-PmrB(LBT) with terminator (BBa_B1006)
PmrA-PmrB two-component system is native to
Salmonella enterica and in it's native state it is responsible for chemotaxis. PmrB is a transmembrane protein with iron binding peptide on it's extracellular loop. When PmrB binds iron (III) iron, it's intracellular domain gains kinase activity and phosphorylates PmrA, which subsequently binds to pmrC promoter and induces expression of chemotaxis CheZ protein. In this part iron binding tag on the extracellular loop was exchanged with a lanthanide binding tag (LBT), to allow PmrA-PmrB two-component system to respond to lanthanide ions.
SENT TO REGISTRY
BBa_K1459005 - PmrA-PmrB(N-term)
This is N-terminal part of PmrA-PmrB two-component system native to
Salmonella enterica. PmrA-PmrB two-component system is native to Salmonella enterica and in it's native state it is responsible for chemotaxis. PmrB is a transmembrane protein with iron binding peptide on it's extracellular loop. When PmrB binds iron (III) iron, it's intracellular domain gains kinase activity and phosphorylates PmrA, which subsequently binds to pmrC promoter and induces expression of chemotaxis CheZ protein. In this part iron binding tag on the extracellular loop was exchanged with a lanthanide binding tag (LBT), to allow PmrA-PmrB two-component system to respond to lanthanide ions. In this part, PmrB protein is truncated just before iron binding tag, which enables one to put any desired tag between two parts of PmrB, to construct a detecting system based on PmrA-PmrB system.
SENT TO REGISTRY
BBa_K1459006 - pmrC
This is pmrC promoter native to
Salmonella enterica. PmrA-PmrB two-component system is native to
Salmonella enterica and in it's native state it is responsible for chemotaxis. PmrB is a transmembrane protein with iron binding peptide on it's extracellular loop. When PmrB binds iron (III) iron, it's intracellular domain gains kinase activity and phosphorylates PmrA, which subsequently binds to pmrC promoter and induces expression of chemotaxis CheZ protein. In this part iron binding tag on the extracellular loop was exchanged with a lanthanide binding tag (LBT), to allow PmrA-PmrB two-component system to respond to lanthanide ions.
BBa_K1459008 - pmrC-GFP-terminator
This is pmrC promoter from
Salmonella enterica, with subsequent GFP and BBa_B1006 terminator. This part is one part of PmrA-PmrB detecting system. Upon phosphorylation by PmrB, PmrA binds to pmrC and induces expression of GFP.
SENT TO REGISTRY
BBa_K1459010 - PmrB(LBT)
PmrB(LBT) is a engineered PmrB gene, in which iron binding tag was replaced by lanthanide binding tag. PmrA-PmrB two-component system is native to
Salmonella enterica and in it's native state it is responsible for chemotaxis. PmrB is a transmembrane protein with iron binding peptide on it's extracellular loop. When PmrB binds iron (III) iron, it's intracellular domain gains kinase activity and phosphorylates PmrA, which subsequently binds to pmrC promoter and induces expression of chemotaxis CheZ protein. In this part iron binding tag on the extracellular loop was exchanged with a lanthanide binding tag (LBT), to allow PmrA-PmrB two-component system to respond to lanthanide ions.
SENT TO REGISTRY
BBa_K1459011 - PmrB(N-term)
PmrA-PmrB two-component system is native to
Salmonella enterica and in it's native state it is responsible for chemotaxis. PmrB is a transmembrane protein with iron binding peptide on it's extracellular loop. When PmrB binds iron (III) iron, it's intracellular domain gains kinase activity and phosphorylates PmrA, which subsequently binds to pmrC promoter and induces expression of chemotaxis CheZ protein.
In this part iron binding tag on the extracellular loop was exchanged with a lanthanide binding tag (LBT), to allow PmrA-PmrB two-component system to respond to lanthanide ions. This part was truncated just before the iron binding tag, and PmrB(N-term) is functionally complementar to PmrB(C-term).
SENT TO REGISTRY
BBa_K1459012 - SENG lanthanide binding tag
This is a DNA sequence coding lanthanide binding tag described in literature. It's literatural dissociation constants are as follows:
K
Tb3+=18 nM
This is the lowest known value of dissociation constant for a Tb
3+, thus making the binding strenght highest amongst known LBTs.
BBa_K1459013 - wSE3 lanthanide binding tag
This is sequence of DNA coding wSE3 lanthanide binding tag. It's dissociation constants are as follows:
K
Tb3+=2000 nM
BBa_K1459014 - Lanthanide Binding Tag
This is DNA sequence coding a lanthanide binding tag. This one is one of the best described LBTs in literature, with dissociation constants following:
K
La3+= 3500 nM
K
Ce3+= 950 nM
K
Nd3+= 270 nM
K
Eu3+= 62 nM
K
Gd3+= 84 nM
K
Tb3+= 57 nM
K
Dy3+= 71 nM
K
Er3+= 78 nM
K
Yb3+= 100 nM
K
Lu3+= 128 nM
BBa_K1459015 - 1L2Y short peptide
This is DNA sequence coding short peptide (PDB 1L2Y) is highly structured in water and could provide a structural foundation for small binding tags, such as we were planning to use it.
Results
Cooperation with other iGEM Teams
During this year’s iGEM we have exchanged with the following teams:
Paris_Bettencourt – we participated in the iGEM newsletter, sending them information about our team, our project and trying to answer other teams questions from the previous newsletter
Toulouse – we sent them 4 of our BioBricks (BBa_K780003, BBa_K780002, BBa_K780001, BBa_K780000)
Groningen – we exchanged our official iGEM abstracts, translated their abstract into Polish and got our abstract translated into Dutch
Paris Saclay - we exchanged our official iGEM abstracts and we translated their into Polish and got our abstract translated into French
ETH Zurich – we filled in a survey about complexity in everyday life
Warwick - we filled in a survey about policy and practices
Valencia Biocampus - we filled in a survey
Medal Criteria
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