Team:LMU-Munich/Weekly Journal

From 2012.igem.org

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BacillusBioBrickBox
BacillusBioBrickBox
<html><a href="https://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks">
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<img src="https://static.igem.org/mediawiki/2012/8/8d/Bacilluss_Intro.png" height=40"/></a></html>
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===3-7 September 2012===
===3-7 September 2012===
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<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
Plates of our spores diluted at 10^-2, 10^-4 and 10^-6 from the germination assay show NO GERMINATION for our triple and quadruple mutants, and plenty of germination for the WT positive control! We will try plating undiluted mutant spores to see if any germination occurs.
Plates of our spores diluted at 10^-2, 10^-4 and 10^-6 from the germination assay show NO GERMINATION for our triple and quadruple mutants, and plenty of germination for the WT positive control! We will try plating undiluted mutant spores to see if any germination occurs.
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<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> A collection of useful tags in Freiburgs standard and with RBS included was cloned into pSB1C3. The tags are: 3xFlag, HA, cMyc, 10xHis and Streptavidin.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> A collection of useful tags in Freiburgs standard and with RBS included was cloned into pSB1C3. The tags are: 3xFlag, HA, cMyc, 10xHis and Streptavidin.
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The genes [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823028 luc+] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823029 mKate2], synthesized by gene art were succesfully cloned into pSB1C3 and sequenced.
The genes [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823028 luc+] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823029 mKate2], synthesized by gene art were succesfully cloned into pSB1C3 and sequenced.
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'''20-24 August 2012'''
'''20-24 August 2012'''
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'''13-17 August 2012'''
'''13-17 August 2012'''
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The xylose-inducible promoter with the according repressor (which has a constitutive promoter, RBS and terminator) P<sub><i>Xyl</> + <i>XylR</i> was cloned into pSB1C3 and sequenced.
The xylose-inducible promoter with the according repressor (which has a constitutive promoter, RBS and terminator) P<sub><i>Xyl</> + <i>XylR</i> was cloned into pSB1C3 and sequenced.
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'''6-10 August 2012'''
'''6-10 August 2012'''
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'''30 July - 3 August 2012'''
'''30 July - 3 August 2012'''
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'''23-27 July 2012'''
'''23-27 July 2012'''
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'''16-20 July 2012'''
'''16-20 July 2012'''
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<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> Cloning of the Anderson promoters J23100, J23102, J23103, J23106 in the reporter vector pSB<sub>''Bs''</sub>1C-''lacZ'' for β-galactosidase assays finished.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> Cloning of the Anderson promoters J23100, J23102, J23103, J23106 in the reporter vector pSB<sub>''Bs''</sub>1C-''lacZ'' for β-galactosidase assays finished.
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'''9-13 July 2012'''
'''9-13 July 2012'''
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> The vectors <b>pSB<sub>Bs</sub>4S-P<sub><i>Xyl</i></sub> </b>, <b>pSB<sub>Bs</sub>1C-<i>lacZ</i> </b> and <b>pSB<sub>Bs</sub>4S </b> were succesfully completed and tested by restriction digest as well as red colony colour.  
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> The vectors <b>pSB<sub>Bs</sub>4S-P<sub><i>Xyl</i></sub> </b>, <b>pSB<sub>Bs</sub>1C-<i>lacZ</i> </b> and <b>pSB<sub>Bs</sub>4S </b> were succesfully completed and tested by restriction digest as well as red colony colour.  
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'''2-6 July 2012'''
'''2-6 July 2012'''
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<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> Cloning of the Anderson promoters J23100, J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118 in pSB1C3 finished.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> Cloning of the Anderson promoters J23100, J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118 in pSB1C3 finished.
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'''25-29 June 2012'''
'''25-29 June 2012'''
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'''18-22 June 2012'''
'''18-22 June 2012'''
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'''11-15 June 2012'''
'''11-15 June 2012'''
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<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
Clean deletions of germination genes ''cwlD'', ''cwlJ'', and ''gerD'' from PCR accomplished. DNA purified and frozen to be later transformed with ''Bacillus''.
Clean deletions of germination genes ''cwlD'', ''cwlJ'', and ''gerD'' from PCR accomplished. DNA purified and frozen to be later transformed with ''Bacillus''.
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'''4-8 June 2012'''
'''4-8 June 2012'''
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'''28-1 June 2012'''
'''28-1 June 2012'''
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'''21-25 May 2012'''
'''21-25 May 2012'''
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<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> The ''Bacillus'' promoters P<sub>''liaG''</sub>, P<sub>''liaI''</sub> and P<sub>''lepA''</sub> are in the ''Bacillus'' reporter vector pSB<sub>''Bs''</sub>3C-''luxABCDE''.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> The ''Bacillus'' promoters P<sub>''liaG''</sub>, P<sub>''liaI''</sub> and P<sub>''lepA''</sub> are in the ''Bacillus'' reporter vector pSB<sub>''Bs''</sub>3C-''luxABCDE''.
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'''14-18 May 2012'''
'''14-18 May 2012'''
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'''7-11 May 2012'''
'''7-11 May 2012'''
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> Cloning of P<sub>''liaG''</sub>, P<sub>''liaI''</sub> and P<sub>''veg''</sub> in vector pSB<sub>''Bs''</sub>1C-''lacZ'' finished.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> Cloning of P<sub>''liaG''</sub>, P<sub>''liaI''</sub> and P<sub>''veg''</sub> in vector pSB<sub>''Bs''</sub>1C-''lacZ'' finished.
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'''30 April-4 Mai 2012'''
'''30 April-4 Mai 2012'''
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'''23-27 April 2012'''
'''23-27 April 2012'''
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> <b>pSB<sub>Bs</sub>3C-<i>luxABCDE</i> </b> with still one PstI site was created with RFP in the multile cloning site to have a vector for promoter measurments. edit: This PstI site was removed later and only that backbone is submitted to the registry.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> <b>pSB<sub>Bs</sub>3C-<i>luxABCDE</i> </b> with still one PstI site was created with RFP in the multile cloning site to have a vector for promoter measurments. edit: This PstI site was removed later and only that backbone is submitted to the registry.
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'''16-20 April 2012'''
'''16-20 April 2012'''
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> The cloning of the reporter vector <b>pSB<sub>Bs</sub>1C-<i>lacZ</i> </b> was finished.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> The cloning of the reporter vector <b>pSB<sub>Bs</sub>1C-<i>lacZ</i> </b> was finished.
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'''9-13 April 2012'''
'''9-13 April 2012'''
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<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
Jara created the single mutant ''cwlD''::kan.
Jara created the single mutant ''cwlD''::kan.
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'''2-6 April 2012'''
'''2-6 April 2012'''
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<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> With <b>pSB<sub>Bs</sub>0K-P<sub><i>spac</i></sub> </b> our first Vector for our <b>B</b>acillus <b>B</b>io<b>B</b>rick <b>B</b>ox was completed.
<html><a><img src="https://static.igem.org/mediawiki/2012/7/78/BacillusBioBrickBox.png" height=40"/></a></html> With <b>pSB<sub>Bs</sub>0K-P<sub><i>spac</i></sub> </b> our first Vector for our <b>B</b>acillus <b>B</b>io<b>B</b>rick <b>B</b>ox was completed.
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'''26-30 March 2012'''
'''26-30 March 2012'''
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<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
Jara tried knocking out ''cwlB'' using the tet resistance cassette. Mutants of ''cwlB''::tet grew very poorly.
Jara tried knocking out ''cwlB'' using the tet resistance cassette. Mutants of ''cwlB''::tet grew very poorly.
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'''19-23 March 2012'''
'''19-23 March 2012'''
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<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
<img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
We decided which genes to knock out for the germination stop. Based on the work of [http://www.ncbi.nlm.nih.gov/pubmed/19554258 J. Kim and W. Schumann (2009)], we decided to knock out genes ''cwlB'', ''gerD'', ''cwlJ'', and ''sleB''. From the research of [http://www.ncbi.nlm.nih.gov/pubmed/11466293 B. Setlow et al (2001)], we also chose ''cwlD''.
We decided which genes to knock out for the germination stop. Based on the work of [http://www.ncbi.nlm.nih.gov/pubmed/19554258 J. Kim and W. Schumann (2009)], we decided to knock out genes ''cwlB'', ''gerD'', ''cwlJ'', and ''sleB''. From the research of [http://www.ncbi.nlm.nih.gov/pubmed/11466293 B. Setlow et al (2001)], we also chose ''cwlD''.
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'''12-16 March 2012'''
'''12-16 March 2012'''
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'''5-9 March 2012'''
'''5-9 March 2012'''
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'''27 February - 2 March 2012'''
'''27 February - 2 March 2012'''
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'''20-24 February 2012'''
'''20-24 February 2012'''

Revision as of 12:21, 11 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

Team-LMU Photo9.jpg

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".

IGEM HQ LMU prize.jpg

[ more news ]

Sporenfreunde
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