Team:LMU-Munich/Bacillus BioBricks

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Latest revision as of 16:41, 26 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 ]

Bacillus BioBrick Box banner.resized WORDS.JPG

B⁴ - 22 core parts for Bacillus subtilis

A major goal of our iGEM project is to introduce B. subtilis as a new chassis for BioBrick-based synthetic biology. For that purpose, we created a toolbox of Bacillus BioBricks to contribute to the registry to make it accessible to many more future iGEM-teams and the entire public microbiology domain! This Bacillus BioBrick Box (B⁴) contains the following Bacillus specific parts:

Vectors	Promoters	Reporters	Affinity tags

pSB_Bs1C pSB_Bs4S pSB_Bs2E pSB_Bs1C-lacZ pSB_Bs3C-luxABCDE pSB_Bs4S-P_xyl pSB_Bs0K-P_spac Sporovector	Anderson P_liaG P_veg P_lepA P_liaI xylR-P_xyl	gfp mkate2 lacZ luc+	Flag His cMyc Strep HA

Since Bacillus subtilis is not an organism commonly used in iGEM, please check out our Introduction to learn more about it.

Bacillus Vectors


We have generated a suite of BioBrick-compatible vectors: three empty insertional backbones with different antibiotic resistances and integration loci, two reporter and two expression vectors.

Bacillus Promoters


To provide a set of promoters of different strength we characterized several promoters in Bacillus subtilis. Both constitutive and inducible promoters are covered.

Bacillus Reporters


We designed and codon-optimized a set of reporters that are commonly used in B. subtilis.

Affinity Tags


We synthesized 5 affinity tags for protein purification. They all are designed in Freiburg standard with an optimized ribosome binding site upstream. We have not yet tested our tags.

Project Navigation


Bacillus Intro	Bacillus BioBrickBox	Sporobeads	Germination STOP

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

@@ Line 1: / Line 1: @@
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+[[File:Bacillus BioBrick Box banner.resized WORDS.JPG|620px|link=]]
-[[Image:BacillusBioBrickBox.png|150px|right]]
-==Bacillus BioBricks==
-We will create a toolbox of <i>Bacillus</i> BioBricks to contribute to the registry.
+[[Image:BacillusBioBrickBox.png|100px|right|link=]]
-'''We would like to introduce ''Bacillus'' to the world of iGEM!!!'''
-[[Image:LMU-Munich_Introduce_Bacillus.jpg|400px|center]]
-This <b>B</b>acillus <b>B</b>io<b>B</b>rick <b>B</b>ox contains compatible vectors and useful promoters.
+=='''B<sup>4</sup>''' - 22 core parts for ''Bacillus subtilis''==
+<br>
+<p align="justify">A major goal of our iGEM project is to [https://2012.igem.org/Team:LMU-Munich/Bacillus_Introduction introduce ''B. subtilis''] as a new chassis for BioBrick-based synthetic biology. For that purpose, we created a toolbox of <i>Bacillus</i> BioBricks to contribute to the registry to make it accessible to many more future iGEM-teams and the entire public microbiology domain! This ''<b>Bacillus'' B</b>io<b>B</b>rick <b>B</b>ox ('''B<sup>4</sup>''') contains the following ''Bacillus'' specific parts:</p>
-Here is a list of all the vectors we cloned and used. Please note that pMAD is not in a BioBrick standard, but it is a useful tool to knock out genes inBacillus subtilis.
-For the use of our vectors, please see our [[Team:LMU-Munich/Lab_Notebook/Protocols|Protocols]] page. A general introduction to Bacillus subtilis and its integrative vectors can be found [[Team:LMU-Munich/Bacillus Introduction|here]]. All vectors have ampicillin as <i>E. coli</i> resistance and RFP as selection marker.
+<div class="box">
+{| width="100%"
-{|
+| style="width:20%"|'''[[Team:LMU-Munich/Bacillus_BioBricks/Vectors|Vectors]]'''
-|-
+| style="width:20%"|'''[[Team:LMU-Munich/Bacillus_BioBricks/Promoters|Promoters]]'''
-|<b>name</b>
+| style="width:20%"|'''[[Team:LMU-Munich/Bacillus_BioBricks/Reporters|Reporters]]'''
-|<b>E. coli res.</b>
+| style="width:20%"|'''[[Team:LMU-Munich/Bacillus_BioBricks/Tags|Affinity tags]]
-|<b>B. subt. res.</b>
-|<b>insertion locus</b>
-|<b>description</b>
-|<b>derived from</b>
-|<b>reference</b>
 |-
-|<b>pSB<sub>Bs</sub>1C-<i>lacZ</i> </b>
+|[[File:LMU Backbone.png|100px|link=Team:LMU-Munich/Bacillus_BioBricks#Bacillus_Vectors|Vectors]]
-|Amp
+|[[File:LMU PromoterIconBC.png|100px|link=Team:LMU-Munich/Bacillus_BioBricks#Bacillus_Promoters]]
-|Cam
+|[[File:LMU Reporter.png|60px|link=Team:LMU-Munich/Bacillus_BioBricks#Bacillus_Reporters]]
-|amyE
+|[[File:Proteinaffinitytagbutton.png|50px|link=Team:LMU-Munich/Bacillus_BioBricks#Affinity_Tags]]
-|with <i>lacZ</i> reporter gene
-|pAC6
-|?
 |-
-|<b>pSB<sub>Bs</sub>4S </b>
+|valign="top"|<font size="2">
-|Amp
+pSB<sub>''Bs''</sub>1C<br>
-|Spec
+pSB<sub>''Bs''</sub>4S<br>
-|thrC
+pSB<sub>''Bs''</sub>2E<br>
-|empty
+pSB<sub>''Bs''</sub>1C-''lac''Z<br>
-|pDG1731
+pSB<sub>''Bs''</sub>3C-''lux''ABCDE<br>
-|?
+pSB<sub>''Bs''</sub>4S-P<sub>''xyl''</sub><br>
-|-
+pSB<sub>''Bs''</sub>0K-P<sub>''spac''</sub><br>
-|<b>pSB<sub>Bs</sub>1C </b>
+'''Sporo'''vector</font>
-|Amp
+|valign="top"|<font size="2">
-|Cam
+Anderson<br>
-|amyE
+P<sub>''liaG''</sub><br>
-|empty
+P<sub>''veg''</sub><br>
-|pDG1662
+P<sub>''lepA''</sub><br>
-|?
+P<sub>''liaI''</sub><br>
-|-
+''xylR''-P<sub>''xyl''</sub></font>
-|<b>pSB<sub>Bs</sub>4S-P<sub><i>Xyl</i></sub> </b>
+|valign="top"|<font size="2">
-|Amp
+''gfp''<br>
-|Spec
+''mkate2''<br>
-|thrC
+''lacZ''<br>
-|with Xylose-inducible promoter
+''luc+''<br></font>
-|pXT
+|valign="top"|<font size="2">
-|?
+Flag<br>His<br>cMyc<br>Strep<br>HA</font>
-|-
-|<b>pSB<sub>Bs</sub>3C-<i>luxABCDE</i> </b>
-|Amp
-|Cam
-|sacA
-|with <i>luxABCDE</i> reporter cassette
-|pAH328
-|?
-|-
-|<b>pSB<sub>Bs</sub>0K-P<sub><i>spac</i></sub> </b>
-|Amp
-|Kan
-|replicative
-|with IPTG inducible promoter
-|pDG148
-|?
-|-
-|<b>pSB<sub>Bs</sub>2E </b>
-|Amp
-|MLS
-|lacA
-|empty
-|pAX01
-|?
 |-
 |}
+</div>
-The number in the vector's name codes for the insertion locus and the following letter for the <i> Bacillus subtilis </i> resistance gene according to the following table:
-{|
-|-
-|<b>number</b>
-|<b>insertion locus</b>
-|<b>letter</b>
-|<b>resistance </b>
-|-
-|<b>0</b>
-|replicative
-|<b>C</b>
-|Chloramphenicol
-|-
-|<b>1</b>
-|amyE (amylase)
-|<b>E</b>
-|MLS (Erythromycin + Lincomycin)
-|-
-|<b>2</b>
-|lacA (laccase ?)
-|<b>K</b>
-|Kanamycin
-|-
-|<b>3</b>
-|sacA (saccase?)
-|<b>S</b>
-|Spectinomycin
-|-
-|<b>4</b>
-|thrC (threonine C)
-|
-|
-|-
-|-
+[[File:NEXT.png|right|80px|link=Team:LMU-Munich/Germination_Stop]]  [[File:BACK.png|left|80px|link=Team:LMU-Munich/Data/differentiation_tour]]
-|}
-The concentrations of the antibiotics and the insertion tests can be found in our protocol section.
-The promoters we submit:
-Pveg, PliaI, PliaG, plepA, Pxyl, Pxyl+XylR
-We also tested the Anderson promoter collection in <i> Bacillus subtilis </i> with pSB<sub>Bs</sub>3C-<i>luxABCDE</i>. The data will be online in our data section soon.
-Furthermore we plan to submit reporter genes optimized for <i> Bacillus subtilis </i>, namely GFP, lacZ, luc and mKate.
-Useful for the registry are also 5 tages in Freiburg Standard (cMyc, 10His, Flag, Strep, HA).
-More details to come soon.
+<p align="justify">Since ''Bacillus subtilis'' is not an organism commonly used in iGEM, please check out our [https://2012.igem.org/Team:LMU-Munich/Bacillus_Introduction Introduction] to learn more about it.</p>
+<div class="box">
+==''Bacillus'' Vectors   [[File:LMU Backbone.png|100px|link=Team:LMU-Munich/Bacillus_BioBricks#Bacillus_Vectors|Vectors]]==
+{| "width=100%" style="text-align:center;" style="align:right"|
+|<p align="justify">We have generated a suite of BioBrick-compatible vectors: three empty insertional backbones with different antibiotic resistances and integration loci, two reporter and two expression vectors.</p>
+|[[File:LMU-Munich-PSBBs1C.png|200px|right|link=Team:LMU-Munich/Bacillus BioBricks/Vectors]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Bacillus BioBricks/Vectors]]
+|}
+</div>
+<div class="box">
+==''Bacillus'' Promoters   [[File:LMU PromoterIconBC.png|100px]]==
+{| "width=100%" style="align:right"|
+|
+<p align="justify">To provide a set of promoters of different strength we characterized several promoters in ''Bacillus subtilis''. Both constitutive and inducible promoters are covered.</p>
+|
+[[File:Promoters overview.png|200px|right|link=Team:LMU-Munich/Bacillus BioBricks/Promoters]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Bacillus BioBricks/Promoters]]
+|}
+</div>
+<div class="box">
+==''Bacillus'' Reporters   [[File:LMU Reporter.png|50px]]==
+{| "width=100%" style="align:right"|
+|
+<p align="justify">We designed and codon-optimized a set of reporters that are commonly used in ''B. subtilis''.</p>
+|
+[[File:LMU GFP.jpg|200px|right|link=Team:LMU-Munich/Bacillus BioBricks/Reporters]]
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Bacillus BioBricks/Reporters]]
+|}
+</div>
+<div class="box">
+==Affinity Tags  [[File:Proteinaffinitytagbutton.png|50px]]==
+{| "width=100%" style="align:right"|
+|
+<p align="justify">We synthesized 5 affinity tags for protein purification. They all are designed in Freiburg standard with an optimized ribosome binding site upstream. We have not yet tested our tags.</p>
+|-
+! colspan="2" |[[File:LMU Arrow purple.png|40px|link=Team:LMU-Munich/Bacillus BioBricks/Tags]]
+|}
+</div>
+<br>
+<br>
+<br>
+<br>
+<div class="box">
+====Project Navigation====
+{| width="100%" align="center" style="text-align:center;"
+|[[File:Bacilluss_Intro.png|100px|link=Team:LMU-Munich/Bacillus_Introduction]]
+|[[File:BacillusBioBrickBox.png|100px|link=Team:LMU-Munich/Bacillus_BioBricks]]
+|[[File:SporeCoat.png|100px|link=Team:LMU-Munich/Spore_Coat_Proteins]]
+|[[File:GerminationSTOP.png|100px|link=Team:LMU-Munich/Germination_Stop]]
+|-
+|[[Team:LMU-Munich/Bacillus_Introduction|<font size="2">'''''Bacillus'''''<BR>Intro</font>]]
+|[[Team:LMU-Munich/Bacillus_BioBricks|<font size="2" face="verdana">'''''Bacillus'''''<BR>'''B'''io'''B'''rick'''B'''ox</font>]]
+|[[Team:LMU-Munich/Spore_Coat_Proteins|<font size="2" face="verdana">'''Sporo'''beads</font>]]
+|[[Team:LMU-Munich/Germination_Stop|<font size="2" face="verdana">'''Germination'''<BR>STOP</font>]]
+|}
+</div>
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