Team:LMU-Munich/Data
From 2012.igem.org
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{{:Team:LMU-Munich/Templates/Page Header|File:Team-LMU_Photo2.jpg}} | {{:Team:LMU-Munich/Templates/Page Header|File:Team-LMU_Photo2.jpg}} | ||
- | [[File: | + | [[File:Data banner.resized WORDS.JPG|620px|link=]] |
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[[Image:BacillusBioBrickBox.png|right|100px|link=https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks]] | [[Image:BacillusBioBrickBox.png|right|100px|link=https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks]] | ||
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====Evaluation of ''B. subtilis'' vectors==== | ====Evaluation of ''B. subtilis'' vectors==== | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">Seven novel vectors were constructed and four already proven to work in ''B. subtilis''</p> | + | |<p align="justify">Seven novel vectors were constructed and four already proven to work in ''B. subtilis''.</p> |
|[[File:LacZ plate.png|right|100px|link=Team:LMU-Munich/Data/Vectors]] | |[[File:LacZ plate.png|right|100px|link=Team:LMU-Munich/Data/Vectors]] | ||
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====Constitutive ''Bacillus'' Promoters==== | ====Constitutive ''Bacillus'' Promoters==== | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">Three novel ''B. subtilis'' promoter BioBricks ( | + | |<p align="justify">Three novel ''B. subtilis'' promoter BioBricks (P<sub>''liaG''</sub>, P<sub>''veg''</sub>, P<sub>''lepA''</sub>) were constructed and functionality verified by luminescence and β-galactosidase assays.</p> |
|[[File:LMU Constitutive preview.jpg|200px|link=Team:LMU-Munich/Data/Constitutive]] | |[[File:LMU Constitutive preview.jpg|200px|link=Team:LMU-Munich/Data/Constitutive]] | ||
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====Inducible ''Bacillus'' Promoters==== | ====Inducible ''Bacillus'' Promoters==== | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">A novel antibiotic-inducible promoter, | + | |<p align="justify">A novel antibiotic-inducible promoter, P<sub>''liaI''</sub>, was constructed and thoroughly evaluated.</p> |
|[[File:Englisch Auswertung PliaI.png|200px|link=Team:LMU-Munich/Data/Inducible]] | |[[File:Englisch Auswertung PliaI.png|200px|link=Team:LMU-Munich/Data/Inducible]] | ||
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===Crust Promoter Evaluation=== | ===Crust Promoter Evaluation=== | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">Three sporulation-dependent promoters ( | + | |<p align="justify">Three sporulation-dependent promoters (P<sub>''cotYZ''</sub>, P<sub>''cotV''</sub>, P<sub>''cgeA''</sub>) were constructed, two of which show the expected behavior.</p> |
|[[File:Promotoren.png|200px|link=Team:LMU-Munich/Data/crustpromoters]] | |[[File:Promotoren.png|200px|link=Team:LMU-Munich/Data/crustpromoters]] | ||
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===GFP-'''Sporo'''bead Evaluation=== | ===GFP-'''Sporo'''bead Evaluation=== | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">They glow! GFP was successfully displayed on the spore crust as | + | |<p align="justify">They glow! GFP was successfully displayed on the spore crust as demonstrated by fluorescence microscopy.</p> |
|[[File:Sporobead Constructs Dada sheet preview.png|200px|link=Team:LMU-Munich/Data/gfp_spore]] | |[[File:Sporobead Constructs Dada sheet preview.png|200px|link=Team:LMU-Munich/Data/gfp_spore]] | ||
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==='''Sporo'''bead Purification=== | ==='''Sporo'''bead Purification=== | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">We compared different ways of purifying ''Bacillus'' spores from vegetative cells: | + | |<p align="justify">We compared different ways of purifying ''Bacillus'' spores from vegetative cells: Lysozyme is the best!</p> |
|[[File:Purification preview.png|200px|link=Team:LMU-Munich/Data/Sporepurification]] | |[[File:Purification preview.png|200px|link=Team:LMU-Munich/Data/Sporepurification]] | ||
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===Knockouts of germination genes === | ===Knockouts of germination genes === | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">We successfully prevented spore germination by multiple gene knockouts</p> | + | |<p align="justify">We successfully prevented spore germination by multiple gene knockouts.</p> |
|[[File:LMU Knockout previewii.jpg|200px|link=Team:LMU-Munich/Data/Knockout]] | |[[File:LMU Knockout previewii.jpg|200px|link=Team:LMU-Munich/Data/Knockout]] | ||
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</div> | </div> | ||
<div class="box"> | <div class="box"> | ||
- | ===Suicide Switch === | + | ==='''Suicide'''Switch === |
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify">Death upon germination: We developed a novel safety strategy as a back-up to knockouts.</p> | + | |<p align="justify">Death upon germination: We developed a novel safety strategy as a back-up to the gene knockouts.</p> |
|[[File:PlatereaderSuicideSwitch1.jpg|200px|link=Team:LMU-Munich/Data/Suicideswitch]] | |[[File:PlatereaderSuicideSwitch1.jpg|200px|link=Team:LMU-Munich/Data/Suicideswitch]] | ||
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===β-galactosidase assay of Inverter with ''lacZα'' === | ===β-galactosidase assay of Inverter with ''lacZα'' === | ||
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
- | |<p align="justify"> | + | |<p align="justify">It works! Invert the output of your promoter of choice.</p> |
|[[File:LMU Inverter graph.png|200px|link=Team:LMU-Munich/Data/Inverter]] | |[[File:LMU Inverter graph.png|200px|link=Team:LMU-Munich/Data/Inverter]] | ||
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{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
|<p align="justify"> | |<p align="justify"> | ||
- | Unexpected restriction sites combined with no activity at all</p> | + | Unexpected restriction sites combined with no activity at all - what a bummer!</p> |
|[[File:LMU cgea preview.jpg|200px|link=Team:LMU-Munich/Data/cgeA]] | |[[File:LMU cgea preview.jpg|200px|link=Team:LMU-Munich/Data/cgeA]] | ||
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{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
|<p align="justify"> | |<p align="justify"> | ||
- | The story of trying to clone a | + | The story of trying to clone a toxin from ''E. coli'' in ''E. coli'': They die, dammit! (...not a surprise, really...) |
</p> | </p> | ||
|[[File:MazFtrashcan.jpg|200px|link=Team:LMU-Munich/Data/MazF]] | |[[File:MazFtrashcan.jpg|200px|link=Team:LMU-Munich/Data/MazF]] | ||
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<div class="box"> | <div class="box"> | ||
- | ===Xylose | + | ===Xylose promoters=== |
{| "width=100%" style="text-align:center;"| | {| "width=100%" style="text-align:center;"| | ||
|<p align="justify"> | |<p align="justify"> | ||
- | All constructs involving | + | All constructs involving xylose-inducible promoters refused cloning or evaluation (sorry, Groningen...!). |
</p> | </p> | ||
|[[File:100px-D-Xylose Keilstrich.png|100px|link=Team:LMU-Munich/Data/Pxyl]] | |[[File:100px-D-Xylose Keilstrich.png|100px|link=Team:LMU-Munich/Data/Pxyl]] |
Latest revision as of 14:31, 22 October 2012
The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".
[ more news ]
BacillusBioBrickBox
Evaluation of B. subtilis vectors
Seven novel vectors were constructed and four already proven to work in B. subtilis. | |
Anderson Promoter Evaluation
Eleven Anderson promoters were measured in B. subtilis and showed relatively weak activity. | |
Constitutive Bacillus Promoters
Three novel B. subtilis promoter BioBricks (PliaG, Pveg, PlepA) were constructed and functionality verified by luminescence and β-galactosidase assays. | |
Inducible Bacillus Promoters
A novel antibiotic-inducible promoter, PliaI, was constructed and thoroughly evaluated. | |
Sporobeads
Crust Promoter Evaluation
Three sporulation-dependent promoters (PcotYZ, PcotV, PcgeA) were constructed, two of which show the expected behavior. | |
GFP-Sporobead Evaluation
They glow! GFP was successfully displayed on the spore crust as demonstrated by fluorescence microscopy. | |
Sporobead Purification
We compared different ways of purifying Bacillus spores from vegetative cells: Lysozyme is the best! | |
GerminationSTOP
Knockouts of germination genes
We successfully prevented spore germination by multiple gene knockouts. | |
SuicideSwitch
Death upon germination: We developed a novel safety strategy as a back-up to the gene knockouts. | |
Inverter
β-galactosidase assay of Inverter with lacZα
It works! Invert the output of your promoter of choice. | |
Trashcan
Things that were a pain in the neck!
MazF
The story of trying to clone a toxin from E. coli in E. coli: They die, dammit! (...not a surprise, really...) | |
Xylose promoters
All constructs involving xylose-inducible promoters refused cloning or evaluation (sorry, Groningen...!). | |
For our big breakthroughs, or to follow specific projects, see the individual project pages: