Team:LMU-Munich/Data

From 2012.igem.org

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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 ]

Data banner.resized WORDS.JPG

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 (P_liaG, P_veg, P_lepA) were constructed and functionality verified by luminescence and β-galactosidase assays.

Inducible Bacillus Promoters


A novel antibiotic-inducible promoter, P_liaI, was constructed and thoroughly evaluated.

Sporobeads

Crust Promoter Evaluation


Three sporulation-dependent promoters (P_cotYZ, P_cotV, P_cgeA) 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!

cgeA and P_cgeA


Unexpected restriction sites combined with no activity at all - what a bummer!

MazF

The story of trying to clone a toxin from E. coli in E. coli: They die, dammit! (...not a surprise, really...)

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...!).

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:

Project Navigation


Bacillus Intro	Bacillus BioBrickBox	Sporobeads	Germination STOP

Retrieved from "http://2012.igem.org/Team:LMU-Munich/Data"

@@ Line 1: / Line 1: @@
 {{:Team:LMU-Munich/Templates/Page Header|File:Team-LMU_Photo2.jpg}}
-[[File:Data_banner.jpg|620px|link=]]
+[[File:Data banner.resized WORDS.JPG|620px|link=]]
+[[Image:BacillusBioBrickBox.png|right|100px|link=https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks]]
-=Data=
-Here you will find all of our project data.
-[[Image:BacillusBioBrickBox.png|right|100px|link=Team:LMU-Munich/Team:LMU-Munich/Bacillus_BioBricks]]
 <br>
 <div class="box" style= "background-color:#e4f1d7">
@@ Line 18: / Line 13: @@
 ====Evaluation of ''B. subtilis'' vectors====
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">Evaluation of ''B. subtilis'' vectors in ''B. subtilis'' with different inserts</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]]
 |-
@@ Line 27: / Line 22: @@
 ====Anderson Promoter Evaluation====
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">Evaluation of the ''Anderson'' library in ''B. subtilis'' with pSB<sub>Bs</sub>3C-''lux'' (luminescence) and pSB<sub>Bs</sub>1C-''lacZ''</p>
+|<p align="justify">Eleven Anderson promoters were measured in ''B. subtilis'' and showed relatively weak activity.</p>
 |[[File:LMU Anderson preview.jpg|200px|link=Team:LMU-Munich/Data/Anderson]]
 |-
@@ Line 36: / Line 31: @@
 ====Constitutive ''Bacillus'' Promoters====
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">Evaluation of constitutive ''B. subtilis'' promoters with pSB<sub>Bs</sub>3C-''lux'' and pSB<sub>Bs</sub>1C-''lacZ''</p>
+|<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]]
 |-
@@ Line 45: / Line 40: @@
 ====Inducible ''Bacillus'' Promoters====
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">Evaluation of inducible ''B. subtilis'' promoters with pSB<sub>Bs</sub>3C-''lux'' and pSB<sub>Bs</sub>1C-''lacZ''</p>
+|<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]]
 |-
@@ Line 59: / Line 54: @@
 <div class="box">
-===Spore purification===
+===Crust Promoter Evaluation===
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">We compared different ways of purifying ''Bacillus'' spores from vegetative cells</p>
+|<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]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/crustpromoters]]
+|}
+</div>
+<div class="box">
+===GFP-'''Sporo'''bead Evaluation===
+{| "width=100%" style="text-align:center;"|
+|<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]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/gfp_spore]]
+|}
+</div>
+<div class="box">
+==='''Sporo'''bead Purification===
+{| "width=100%" style="text-align:center;"|
+|<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]]
 |-
@@ Line 67: / Line 82: @@
 |}
 </div>
 [[File:GerminationSTOP.png|90px|right|link=Team:LMU-Munich/Germination_Stop]]
@@ Line 77: / Line 94: @@
 ===Knockouts of germination genes ===
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">Germination assy of ''Bacillus'' strains with multiple gene knockouts</p>
+|<p align="justify">We successfully prevented spore germination by multiple gene knockouts.</p>
-|[[File:LMU Knockout preview.jpg|200px|link=Team:LMU-Munich/Data/Knockout]]
+|[[File:LMU Knockout previewii.jpg|200px|link=Team:LMU-Munich/Data/Knockout]]
 |-
 ! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/Knockout]]
 |}
 </div>
+<div class="box">
+==='''Suicide'''Switch ===
+{| "width=100%" style="text-align:center;"|
+|<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]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/Suicideswitch]]
+|}
+</div>
+[[File:LMU-Munich-Invertersign.png|90px|right|link=Team:LMU-Munich/Inverter]]
+<br>
 <div class="box" style= "background-color:#e4f1d7">
+===Inverter===
 </div>
+<div class="box">
+===β-galactosidase assay of Inverter with ''lacZα'' ===
+{| "width=100%" style="text-align:center;"|
+|<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]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/Inverter]]
+|}
+</div>
+<div class="box" style= "background-color:#e4f1d7">
+==='''Trash'''can===
+Things that were a pain in the ''neck''!
+</div>
+<div class="box">
+===''cge''A and P<sub>''cge''A</sub>===
+{| "width=100%" style="text-align:center;"|
+|<p align="justify">
+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]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/cgeA]]
+|}
+</div>
+<div class="box">
+===MazF===
+{| "width=100%" style="text-align:center;"|
+|<p align="justify">
+The story of trying to clone a toxin from ''E. coli'' in ''E. coli'': They die, dammit! (...not a surprise, really...)
+</p>
+|[[File:MazFtrashcan.jpg|200px|link=Team:LMU-Munich/Data/MazF]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/MazF]]
+|}
+</div>
+<div class="box">
+===Xylose promoters===
+{| "width=100%" style="text-align:center;"|
+|<p align="justify">
+All constructs involving xylose-inducible promoters refused cloning or evaluation (sorry, Groningen...!).
+</p>
+|[[File:100px-D-Xylose Keilstrich.png|100px|link=Team:LMU-Munich/Data/Pxyl]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Data/Pxyl]]
+|}
+</div>
 <p></p>