Difference between revisions of "Team:INSA-UPS France/Parts"
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<p> | <p> | ||
| − | + | <b>To learn more: </b> see our <a href="https://2017.igem.org/Team:INSA-UPS_France/Design">design</a>, and our<a href="https://2017.igem.org/Team:INSA-UPS_France/Results"> results</a> | |
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<p> | <p> | ||
| − | <b>see our <a href="https://2017.igem.org/Team:INSA-UPS_France/Design">design</a>, and our<a href="https://2017.igem.org/Team:INSA-UPS_France/Results"> results</a> | + | <b> <b>To learn more: </b> see our <a href="https://2017.igem.org/Team:INSA-UPS_France/Design">design</a>, and our<a href="https://2017.igem.org/Team:INSA-UPS_France/Results"> results</a> |
| − | + | ||
<h2><i>Vibrio harveyi</i> complete gene circuit: </h2> | <h2><i>Vibrio harveyi</i> complete gene circuit: </h2> | ||
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<p> | <p> | ||
| − | <b>To learn more: </b> </html><partinfo>BBa_K2278021</partinfo><html>,<a href="https://2017.igem.org/Team:INSA-UPS_France/Results">results</a> | + | <b>To learn more: </b> see our</html><partinfo>BBa_K2278021</partinfo><html>,and our <a href="https://2017.igem.org/Team:INSA-UPS_France/Results">results</a> |
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<figure> | <figure> | ||
| − | <img src="https://static.igem.org/mediawiki/parts/ | + | <img src="https://static.igem.org/mediawiki/parts/c/cc/T--INSA-UPS_France--pAOXI-leucrocinI.png" alt=""> |
<figcaption> | <figcaption> | ||
fig: <i>Pichia pastoris</i> Leucrocin I AMP generator | fig: <i>Pichia pastoris</i> Leucrocin I AMP generator | ||
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<figure> | <figure> | ||
| − | <img src="https://static.igem.org/mediawiki/parts/ | + | <img src="https://static.igem.org/mediawiki/parts/4/46/T--INSA-UPS_France--pAOXI-cOT2.png |
" alt=""> | " alt=""> | ||
<figcaption> | <figcaption> | ||
Revision as of 00:32, 24 October 2017
BBa_J04450 was tested in the Vibrio harveyi background. BBa_J04450 biobrick was cloned in a broad host range plasmid (pBBR1MCS-4) and conjugated into Vibrio harveyi to demonstrate the production of RFP in this chassis.
To learn more: <partinfo>BBa_J04450</partinfo>, results
The pGAP promoter was used as a constitutive promoter. The promoter was characterized by RT-qPCR using d-ny15 as reporter gene.
To learn more: <partinfo>BBa_K1800001</partinfo>, see our results
The α-factor sequence contains a kozak region and a signal sequence to secrete the produced peptides. The functionality of the signal factor was investigated by demonstrating that AMP are present in the supernatant through a toxicity assay.
To learn more: <partinfo>BBa_K180001</partinfo>,results
This part contains the first half of Vibrio harveyi quorum CqsS receptor which is able to detect CAI-1 family molecule. The receptor is driven by a constitutive promoter.
This part has been designed due to IDT gBlocks synthesis requirements. The ordered gBlocks size should not exceed 3kb (the total length of the Vibrio harveyi circuit is 6929 bp). The XhoI restriction site naturally present in the gene was used reassemble the cqsS* sequence. Therefore, it was impossible to use pSB1C3 vector because of the presence of a XhoI restriction on the vector. This part was then subcloned in pBR322.
To learn more: see our design, and our results
This part contains the second half of Vibrio harveyi quorum CqsS* receptor which is able to detect CAI-1 family molecule. It also contains the tetR repressor under the control of pQRR4 promoter.
This part has been designed due to IDT gBlocks synthesis requirements. The ordered gBlocks size should not exceed 3kb (the total length of the Vibrio harveyi circuit is 6929 bp). The ApaI (not present on pSB1C3 vector) and XhoI restriction (naturally present in the cqsS* gene) were used to reassemble the synthetic circuit.
To learn more: see our design, and our results
This part includes the complete cqsS* gene driven by a constitutive promoter and the tetracyclin repressor under the control of pQRR4 promoter which activation depends on CqsS* detection of quorum sensing inducers.
Vh1+2 is the assembly of Vh2 in Vh1. This part was cloned in pBR322.
To learn more: see our design, and our results
This part includes the complete cqsS gene driven by a constitutive promoter and the tetracycline repressor (TetR) under the control of pQRR4 promoter which activation depends on CqsS* recognition of quorum sensing inducers. The als gene, implied in the diacetyl production pathway, is under the control of pTet which can be repressed by TetR
This part is the assemblage of Vh1, Vh2 and BBa_K2278001. It allows to produce diacetyl is produced upon detection of V. cholerae quorum sensing molecule CAI-1.
The als gene is flanked with two restriction enzyme sites to clone a reporter gene such as rfp.
The assembly of this part was not released due to technical and time issues.
To learn more: see our design, and our results
This system allowed us to constitutively express BBa_K2278021 with a yeast promoter. This system was used to produce AMP at the highest concentration
The pGAP promoter ensures genome recombination in Pichia pastoris genome. The α-factor sequence contains a RBS and a signal sequence to secrete the produced peptides
The restriction enzyme sites allow to extract individually each components of the plasmid. This construction was cloned in pPICZalpha yeast vector.
To learn more: see our<partinfo>BBa_K2278021</partinfo>,and our results
This system allowed us to constitutively express the BBa_K2278022 with a yeast promoter. This system was used to produce AMP at the highest concentration
The pAOXI promoter makes genome recombination easier in Pichia pastoris genome. The α-factor sequence contains a RBS and a signal sequence to secrete the produced peptides
The restriction enzyme sites allow to extract individually each components of the plasmid.
This construction was cloned in pPICZalpha yeast vector.
To learn more: <partinfo>BBa_K2278022</partinfo>,results
This system allowed us to constitutively express BBa_K2278023 with a yeast promoter. This system was used to produce AMP at the highest concentration
The pAOXI promoter makes genome recombination easier in Pichia pastoris genome. The α-factor sequence contains a RBS and a signal sequence to secrete the produced peptides
The restriction enzyme sites allow to extract individually each components of the plasmid.
This construction was cloned in pPICZalpha yeast vector.
To learn more: <partinfo>BBa_K2278023</partinfo>,results
*************************************************
This system allowed us to constitutively express Odr10 receptor to detect diacetyl at any time. Odr10 pathway leads to the activation of pFUS promoter in case of diacetyl detection, hence the AMP are produced.
The α-factor signal and coT2 gene are flanked with two restriction enzyme sites to clone a reporter gene such as RFP.
This construction was cloned in pPICZalpha yeast vector.
To learn more: <partinfo>BBa_K2278023</partinfo>,results
This system allowed us to constitutively express BBa_K2278023 with a yeast promoter. This system was used to produce AMP at the highest concentration
The pGAP promoter makes genome recombination easier in Pichia pastoris genome. The α-factor sequence contains a RBS and a signal sequence to secrete the produced peptides
The restriction enzyme sites allow to extract individually each components of the plasmid.
This construction was cloned in pPICZalpha yeast vector.
To learn more: <partinfo>BBa_K2278023</partinfo>,design
----------------------------------------------------------
iGEM UPS-INSA Toulouse 2017
Parts
New parts submitted to the registry
Name
Function
Type
Part
State
<partinfo>BBa_K2278001</partinfo>
QS molecule generator
basic
working
<partinfo>BBa_K2278002</partinfo>
QS molecule generator
basic
not released
<partinfo>BBa_K2278011</partinfo>
Diacetyl generator
basic
issues
<partinfo>BBa_K2278021</partinfo>
D-NY15 AMP generator
basic
Working
<partinfo>BBa_K2278022</partinfo>
Leucrocin I AMP generator
basic
unsuccessful
<partinfo>BBa_K2278023</partinfo>/td>
coT2 AMP generator
basic
unsuccessful
Existing Parts we have contributed to characterized
<partinfo>BBa_J04450</partinfo>: RFP coding device
<partinfo>BBa_K431009</partinfo>: glyceraldehyde 3-phosphate dehydrogenase promoter (pGAP)
<partinfo>BBa_K1800001</partinfo>: alpha factor secretion signal
Parts used in our project but not submitted to the registry
Vh1: First part of Vibrio harveyi gene circuit
Vh 2 : Second part of Vibrio harveyi gene circuit.
Vh 1+2: Vibrio harveyi invertor system
Vibrio harveyi complete gene circuit:
Pichia pastoris D-NY15 AMP generator:
Pichia pastoris Leucrocin I AMP generator
Pichia pastoris cOT2 AMP generator:
Pichia pastoris Pichia pastoris complete module : Pichia pastoris complete module :
Pichia pastoris Pichia pastoris complete Respond/Effector Module :
Parts used in our project but not submitted to the registry
