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=]]
-=Data=
-Here you will find all of our project data.
 [[Image:BacillusBioBrickBox.png|right|100px|link=https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks]]
@@ 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''' promoters in ''B. subtilis'' with the ''lux'' operon (luminescence) and the reporter ''lacZ'' (β-galactosidase assays).</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 the constitutive ''B. subtilis'' promoters '''P<sub>''liaG''</sub>''', '''P<sub>''veg''</sub>''' and '''P<sub>''lepA''</sub>''' with the ''lux'' operon (luminescence) and the reporter ''lacZ'' (β-galactosidase assays).</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 the inducible ''B. subtilis'' promoter '''P<sub>''liaI''</sub>''' with the ''lux'' operon (luminescence) and the reporter ''lacZ'' (β-galactosidase assays).</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">
-===Promoter evaluation===
+===Crust Promoter Evaluation===
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">We evaluated the three promoters for the crust proteins</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]]
 |-
@@ Line 69: / Line 64: @@
 <div class="box">
-===GFP-Sporobead Evaluation===
+===GFP-'''Sporo'''bead Evaluation===
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">We fused a GFP to the spore crust with five different constructs in two different genetic ''B. subtilis'' strains and analysed the strength with fluorescence microscopy</p>
+|<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]]
 |-
@@ Line 79: / Line 74: @@
 <div class="box">
-===Spore purification===
+==='''Sporo'''bead Purification===
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">We compared different ways of purifying ''Bacillus'' spores from vegetative cells</p>
+|<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 99: / 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 previewii.jpg|200px|link=Team:LMU-Munich/Data/Knockout]]
 |-
@@ Line 106: / Line 101: @@
 </div>
 <div class="box">
-===Suicide Switch ===
+==='''Suicide'''Switch ===
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">Platereader assay of <b>Suicide</b> switch with lux in the place of MazF with PspoIVB</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]]
 |-
@@ Line 122: / Line 117: @@
 <div class="box">
-===ONPG assay of Inverter with lacZalpha ===
+===β-galactosidase assay of Inverter with ''lacZα'' ===
 {| "width=100%" style="text-align:center;"|
-|<p align="justify">ONPG assay of Inverter with lacZalpha dependent on Arabinose concentrations</p>
+|<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]]
 |-
@@ Line 141: / Line 136: @@
 {| "width=100%" style="text-align:center;"|
 |<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 Knockout preview.jpg|200px|link=Team:LMU-Munich/Data/cgeA]]
+|[[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]]
@@ Line 152: / Line 147: @@
 {| "width=100%" style="text-align:center;"|
 |<p align="justify">
-The story of trying to clone a toxine from ''E. coli'' in ''E. coli''
+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]]
@@ Line 161: / Line 156: @@
 <div class="box">
-===Xylose-promoters===
+===Xylose promoters===
 {| "width=100%" style="text-align:center;"|
 |<p align="justify">
-All constructs involving Xylose-inducible promoter refused cloning or evaluation.
+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]]