Team:LMU-Munich/Bacillus BioBricks

From 2012.igem.org

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<p align="justify">For the use of our vectors please see our [[Team:LMU-Munich/Lab_Notebook/Protocols|Protocols]] page. A general introduction to [[Team:LMU-Munich/Bacillus Introduction |''Bacillus subtilis'']] and its integrative vectors can be found [[Team:LMU-Munich/Bacillus_BioBricks/integration|here]]. All vectors have ampicillin as <i>Escherichia coli</i> resistance and RFP in the multiple cloning site as selection marker.</p>
<p align="justify">For the use of our vectors please see our [[Team:LMU-Munich/Lab_Notebook/Protocols|Protocols]] page. A general introduction to [[Team:LMU-Munich/Bacillus Introduction |''Bacillus subtilis'']] and its integrative vectors can be found [[Team:LMU-Munich/Bacillus_BioBricks/integration|here]]. All vectors have ampicillin as <i>Escherichia coli</i> resistance and RFP in the multiple cloning site as selection marker.</p>
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====Constitutive promoters from ''B. subtilis'' as novel BioBricks====
====Constitutive promoters from ''B. subtilis'' as novel BioBricks====
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<p align="justify">The second group of promoters are a set of constitutive promoters from ''B. subtilis'' that we have added to the Registry. We evaluated the promoters [http://partsregistry.org/Part:BBa_K823000 P<sub>''liaG''</sub>], [http://partsregistry.org/Part:BBa_K823003 P<sub>''veg''</sub>] and [http://partsregistry.org/Part:BBa_K823002 P<sub>''lepA''</sub>] using the ''lux'' operon, as well as the ''lacZ'' gene as reporters.</p>
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<p align="justify">The second group of promoters are a set of constitutive promoters from ''B. subtilis'' that we have added to the Registry. We evaluated the promoters [http://partsregistry.org/Part:BBa_K823000 P<sub>''liaG''</sub>], [http://partsregistry.org/Part:BBa_K823003 P<sub>''veg''</sub>] and [http://partsregistry.org/Part:BBa_K823002 P<sub>''lepA''</sub>] using the ''lux'' operon as well as the ''lacZ'' gene as reporters.</p>
*'''P<sub>''liaG''</sub>'''    ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K823000 BioBrick:BBa_K823000])
*'''P<sub>''liaG''</sub>'''    ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K823000 BioBrick:BBa_K823000])
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<p align="justify">P<sub>''liaG''</sub> is a constitutive promoter from ''B. subtilis''. It is responsible for the transcription of the last four genes of the ''liaIHGFSR'' locus and hence for the production of the components of the LiaRS system, which is important for the detection of cell wall antibiotics [http://www.ncbi.nlm.nih.gov/pubmed?term=Journal%20of%20Bacteriology%2C%20188%20%2814%29%3A%205153%E2%80%935166: (Jordan ''et al.'', 2006)]. P<sub>''liaG''</sub> was evaluated with the ''lux'' operon (see [https://2012.igem.org/Team:LMU-Munich/Data/Constitutive Data constitutive promoters] [[File:Lux operon.png|100px|link=https://2012.igem.org/Team:LMU-Munich/Data/Constitutive]]) as well as the ''lacZ'' (see [https://2012.igem.org/Team:LMU-Munich/Data/Constitutive Data constitutive promoters] [[File:LacZ.png|50px|link=https://2012.igem.org/Team:LMU-Munich/Data/Constitutive]]) as reporter. This promoter showed a much higher activity than the Anderson promoters, but was still relatively weak in comparison to other evaluated ''Bacillus'' promoters. </p>
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<p align="justify">P<sub>''liaG''</sub> is a constitutive promoter from ''B. subtilis''. It is responsible for the transcription of the last four genes of the ''liaIHGFSR'' locus and hence for the production of the components of the LiaRS system, which is important for the detection of cell wall antibiotics [http://www.ncbi.nlm.nih.gov/pubmed?term=Journal%20of%20Bacteriology%2C%20188%20%2814%29%3A%205153%E2%80%935166: (Jordan ''et al.'', 2006)]. P<sub>''liaG''</sub> was evaluated with the ''lux'' operon (see [https://2012.igem.org/Team:LMU-Munich/Data/Constitutive#Luminescence_measurements Data constitutive promoters] [[File:Lux operon.png|100px|link=https://2012.igem.org/Team:LMU-Munich/Data/Constitutive#Luminescence_measurements]]) as well as the ''lacZ'' (see [https://2012.igem.org/Team:LMU-Munich/Data/Constitutive Data constitutive promoters] [[File:LacZ.png|50px|link=https://2012.igem.org/Team:LMU-Munich/Data/Constitutive]]) as reporter. This promoter showed a much higher activity than the Anderson promoters, but was still relatively weak in comparison to other evaluated ''Bacillus'' promoters. </p>
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*'''P<sub>''lepA''</sub>'''    ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K823002 BioBrick:BBa_K823002])
*'''P<sub>''lepA''</sub>'''    ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K823002 BioBrick:BBa_K823002])
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<p align="justify"> P<sub>''lepA''</sub> is a constitutive promoter that is important for the transcription of the bicistronic operon. One of the expressed proteins is the protein P<sub>''lepA''</sub> [http://www.ncbi.nlm.nih.gov/pubmed?term=Microbiology%2C%20142%3A%201641%E2%80%931649: (Homuth ''et al.'', 1996)]. P<sub>''lepA''</sub> plays an important role during translation as it can move the mRNA-tRNA complex one step back in the ribosome which is expected to improve the fidelity of translation [http://www.ncbi.nlm.nih.gov/pubmed?term=Cell%2C%20127%20%284%29%3A%20721%E2%80%93733: (Qin ''et al.'', 2006)]. This protein was evaluated with the ''lux'' operon as a reporter (see [https://2012.igem.org/Team:LMU-Munich/Data/Constitutive Data constitutive promoters] [[File:Lux operon.png|100px|link=https://2012.igem.org/Team:LMU-Munich/Data/Constitutive]]). The activity of this promoter is between the activity of the strongest ''Bacillus'' promoter P<sub>''veg''</sub> and the weak P<sub>''liaG''</sub>. </p>
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<p align="justify"> P<sub>''lepA''</sub> is a constitutive promoter that is important for the transcription of the bicistronic operon. One of the expressed proteins is the protein P<sub>''lepA''</sub> [http://www.ncbi.nlm.nih.gov/pubmed?term=Microbiology%2C%20142%3A%201641%E2%80%931649: (Homuth ''et al.'', 1996)]. P<sub>''lepA''</sub> plays an important role during translation as it can move the mRNA-tRNA complex one step back in the ribosome which is expected to improve the fidelity of translation [http://www.ncbi.nlm.nih.gov/pubmed?term=Cell%2C%20127%20%284%29%3A%20721%E2%80%93733: (Qin ''et al.'', 2006)]. This protein was evaluated with the ''lux'' operon as a reporter (see [https://2012.igem.org/Team:LMU-Munich/Data/Constitutive#Luminescence_measurements Data constitutive promoters] [[File:Lux operon.png|100px|link=https://2012.igem.org/Team:LMU-Munich/Data/Constitutive#Luminescence_measurements]]). The activity of this promoter is between the activity of the strongest ''Bacillus'' promoter P<sub>''veg''</sub> and the weak P<sub>''liaG''</sub>. </p>

Revision as of 22:58, 26 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

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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".

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