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Team:Bielefeld-Germany/Results/Submitted
From 2010.igem.org
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| - | = | + | = Submitted BioBricks = |
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| + | We have submitted the following BioBricks yet: | ||
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| + | ===''virA'' receptor=== | ||
| + | The VirA receptor is used by ''A. tumefaciens'' to detect acetosyringone and other phenolic substances which are secreted by plants after injury. In presence of these substances VirA phosphorylates VirG, a response regulator which activates ''vir'' promotors. These promotors control genes which are used for infecting the injured plant. | ||
| + | Actually we wanted to use the ''virA'' receptor already existing in the partsregistry (<partinfo>K238008</partinfo>). But due to some problems (compare results in BioBricks/tested) we decided to isolate the ''virA'' gene from the TI-plasmid of ''A. tumefaciens'' C58 ourselves and bring it into a BioBrick compatible form. We removed an illegal ''PstI'' restriction site in the ''virA'' gene by site-directed mutagenesis. | ||
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| + | You can find this BioBrick here: <partinfo>K389001</partinfo> | ||
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| + | You can find this BioBrick under the control of a constitutive promotor (<partinfo>J23110</partinfo>) here: <partinfo>K389010</partinfo>. | ||
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| + | ===Mutated ''virG''=== | ||
| + | Phosphorylated VirG binds to ''vir'' promotors and activates them. VirG is activated by the acetosyringone receptor VirA. | ||
| + | This version of VirG activates ''vir'' promotors in ''Escherichia coli'' without the ''rpoA''-gene from ''Agrobacterium tumefaciens''. For this reason the point mutations G56V and I77V are brought into the molecule (compare YC Jung ''et al.'', 2004). Because this BioBrick is synthesized (Mr.Gene GmbH), codon usage is optimized for ''E. coli'' and illegal restriction sites were removed. When you use this ''virG'' gene in a ''virA/G'' signaling system you do not need <partinfo>K238010</partinfo> anymore to get the system working in ''E. coli''. | ||
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| + | You can find this BioBrick here: <partinfo>K389002</partinfo> | ||
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| + | ===''virB''-promoter=== | ||
| + | ''Vir''-promoters from ''A. tumefaciens'' are induced by phosphorylated VirG response regulators and control genes for infecting plants in their natural host. They are part of the VirA/G signal transduction system. | ||
| + | We wanted to use the ''vir''-promotor from the partsregistry (<partinfo>K238011</partinfo>) but the same problems occurred like with the use of the ''virA'' receptor from the partsregistry. So we also have to create a new ''vir''-promoter BioBrick (again from TI-plasmid of ''A. tumefaciens'' C58). | ||
| + | |||
| + | You can find this BioBrick here: <partinfo>K389003</partinfo> | ||
Revision as of 09:29, 21 October 2010
Contents |
Submitted BioBricks
We have submitted the following BioBricks yet:
virA receptor
The VirA receptor is used by A. tumefaciens to detect acetosyringone and other phenolic substances which are secreted by plants after injury. In presence of these substances VirA phosphorylates VirG, a response regulator which activates vir promotors. These promotors control genes which are used for infecting the injured plant. Actually we wanted to use the virA receptor already existing in the partsregistry (<partinfo>K238008</partinfo>). But due to some problems (compare results in BioBricks/tested) we decided to isolate the virA gene from the TI-plasmid of A. tumefaciens C58 ourselves and bring it into a BioBrick compatible form. We removed an illegal PstI restriction site in the virA gene by site-directed mutagenesis.
You can find this BioBrick here: <partinfo>K389001</partinfo>
You can find this BioBrick under the control of a constitutive promotor (<partinfo>J23110</partinfo>) here: <partinfo>K389010</partinfo>.
Mutated virG
Phosphorylated VirG binds to vir promotors and activates them. VirG is activated by the acetosyringone receptor VirA. This version of VirG activates vir promotors in Escherichia coli without the rpoA-gene from Agrobacterium tumefaciens. For this reason the point mutations G56V and I77V are brought into the molecule (compare YC Jung et al., 2004). Because this BioBrick is synthesized (Mr.Gene GmbH), codon usage is optimized for E. coli and illegal restriction sites were removed. When you use this virG gene in a virA/G signaling system you do not need <partinfo>K238010</partinfo> anymore to get the system working in E. coli.
You can find this BioBrick here: <partinfo>K389002</partinfo>
virB-promoter
Vir-promoters from A. tumefaciens are induced by phosphorylated VirG response regulators and control genes for infecting plants in their natural host. They are part of the VirA/G signal transduction system. We wanted to use the vir-promotor from the partsregistry (<partinfo>K238011</partinfo>) but the same problems occurred like with the use of the virA receptor from the partsregistry. So we also have to create a new vir-promoter BioBrick (again from TI-plasmid of A. tumefaciens C58).
You can find this BioBrick here: <partinfo>K389003</partinfo>
