Team:LMU-Munich/Germination Stop

From 2012.igem.org

(Difference between revisions)
Line 156: Line 156:
<p align="justify">We take advantage of an alternative σ factor and a promoter regulated by it both not present in ''B. subtilis'' and therefore not recognised by any other σ factor of ''B. subtilis''.</p>
<p align="justify">We take advantage of an alternative σ factor and a promoter regulated by it both not present in ''B. subtilis'' and therefore not recognised by any other σ factor of ''B. subtilis''.</p>
<p align="justify">The alternative sigma factor [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823043 ecf41<sub>Bli aa1-204</sub>] ([http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3426412/ Wecke T., Mascher T. 2012]), which derives from ''B. lincheniformes'' a truncated version constitutively on is synthetically linked to a sigma G regulated promotor responding quite late to σ<sup>G</sup>  ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K823048 P<sub>''spoIVB''</sub>] ([http://www.ncbi.nlm.nih.gov/pubmed/15699190 Steil L., Völker U. et al. 2005]) responding strongly or [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823042 P<sub>sspK</sub>] ([http://www.ncbi.nlm.nih.gov/pubmed/15699190 Steil L., Völker U. et al. 2005]) responding weakly). σ<sup>G</sup> is the last σ factor activated in the forespore. Consequently ecf41<sub>Bli aa1-204</sub> is produced quite late in the forespore. ecf41<sub>Bli aa1-204</sub> then activates the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823041 P<sub>''ydfG''</sub>] ([http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3426412/ Wecke T., Mascher T. 2012]) promotor, which is the only target promoter of ecf41<sub>Bli aa1-204</sub>. This promoter is fused to [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823044 MazF] ([http://jcs.biologists.org/content/118/19/4327.abstract Engelberg-Kulka H.,Amitai S. 2005]). Activation of P<sub>''ydfG''</sub> therefore leads to the expression of MazF, a bacterial toxin from ''E.coli'' degrading mRNA. <br></p>
<p align="justify">The alternative sigma factor [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823043 ecf41<sub>Bli aa1-204</sub>] ([http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3426412/ Wecke T., Mascher T. 2012]), which derives from ''B. lincheniformes'' a truncated version constitutively on is synthetically linked to a sigma G regulated promotor responding quite late to σ<sup>G</sup>  ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K823048 P<sub>''spoIVB''</sub>] ([http://www.ncbi.nlm.nih.gov/pubmed/15699190 Steil L., Völker U. et al. 2005]) responding strongly or [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823042 P<sub>sspK</sub>] ([http://www.ncbi.nlm.nih.gov/pubmed/15699190 Steil L., Völker U. et al. 2005]) responding weakly). σ<sup>G</sup> is the last σ factor activated in the forespore. Consequently ecf41<sub>Bli aa1-204</sub> is produced quite late in the forespore. ecf41<sub>Bli aa1-204</sub> then activates the [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823041 P<sub>''ydfG''</sub>] ([http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3426412/ Wecke T., Mascher T. 2012]) promotor, which is the only target promoter of ecf41<sub>Bli aa1-204</sub>. This promoter is fused to [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823044 MazF] ([http://jcs.biologists.org/content/118/19/4327.abstract Engelberg-Kulka H.,Amitai S. 2005]). Activation of P<sub>''ydfG''</sub> therefore leads to the expression of MazF, a bacterial toxin from ''E.coli'' degrading mRNA. <br></p>
-
<p align="justify">The timecourse is as follows: Sporulation of the <b>Sporo</b>beads leads to ecf41<sub>Bli aa1-204</sub>, through the activation of either P<sub>''spoIVB''</sub> or P<sub>sspK</sub>. This then would lead to the expression of MazF over the activation of P<sub>''ydfG''</sub>. This is either not toxic to the <b>Sporo</b>beads, as they do not rely on translation, or not produced any more by the <b>Sporo</b>beads as they are mature spores. After Germination MazF will then be produced for sure and kill the vegetative cell<br></p>
+
<p align="justify">The timecourse is as follows: Sporulation of the <b>Sporo</b>beads leads to ecf41<sub>Bli aa1-204</sub>, through the activation of either P<sub>''spoIVB''</sub> or P<sub>sspK</sub>. This then would lead to the expression of MazF over the activation of P<sub>''ydfG''</sub>. This is either not toxic to the <b>Sporo</b>beads, as they do not rely on translation, or not produced any more by the <b>Sporo</b>beads as they are mature spores. After germination, MazF will then be produced for sure and kill the vegetative cell.<br></p>
<p align="justify">
<p align="justify">
See what [https://2012.igem.org/Team:LMU-Munich/Data/Suicideswitch Data] we get when measuring a module with ''luxABCDE'' instead of MazF.
See what [https://2012.igem.org/Team:LMU-Munich/Data/Suicideswitch Data] we get when measuring a module with ''luxABCDE'' instead of MazF.

Revision as of 17:40, 26 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

Team-LMU streaked plate.resized.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