Team:LMU-Munich/Germination Stop

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Revision as of 12:47, 4 September 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".

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Germination Stop

The goal of this project is to remove the germination capability, while keeping the necessary structural functions of the spores intact.

Based on the work of [http://www.ncbi.nlm.nih.gov/pubmed/19554258 J. Kim and W. Schumann (2009)] and [http://www.ncbi.nlm.nih.gov/pubmed/11466293 B. Setlow et al (2001)], we decided to knock out genes cwlJ, sleB, cwlB,gerD, and cwlD. The genes cwlJ and sleB code for lytic enzymes which are active in the process of germination. In the work of Setlow et al, when cwlJ and sleB were knocked out together, germination frequency was reduced by 5 orders of magnitude. When these genes are knocked out, we hope to yield a B. subtilis strain which produces spores incapable of germination.

Two methods are being employed to knock out germination: resistance cassette (RC) knockouts and clean deletions. Single RC knockouts were created, and have then been combined to create multiple knockouts.

This will be checked with germination assays.

Further, we will investigate the possibility of using a redundant toxin system to immediately kill off any spores which somehow germinate.

Project Navigation

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Bacillus Intro	Bacillus BioBrickBOX	SporeCoat FusionProteins	Germination STOP

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

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 ==Germination Stop==
-The goal of our project '''BEAD'''zillus is the production of ''Bacillus subtilis'' spores which display novel proteins on their outer coat. These novel proteins could be used in a variety of filtration systems, such as binding viruses out of blood, metals out of drinking water, or plastic molecules out of the marine environment. However, before introducing our filtering-spore “beads” into the environment, we must ensure that we are not simultaneously introducing a GMO capable of reproducing on its own.
+The goal of this project is to remove the germination capability, while keeping the necessary structural functions of the spores intact.
-Spores of ''B. subtilis'' are capable of surviving radiation, high heat, freezing, high pressure, and a variety of chemicals. When eventually presented with the right conditions, they can then germinate into vegetative ''B. subtilis'' cells. Thus, simply exposing the spores to harsh conditions does not guarantee that they will never germinate.
+Based on the work of [http://www.ncbi.nlm.nih.gov/pubmed/19554258 J. Kim and W. Schumann (2009)] and [http://www.ncbi.nlm.nih.gov/pubmed/11466293 B. Setlow et al (2001)], we decided to knock out genes ''cwlJ'', ''sleB'', ''cwlB'',''gerD'', and ''cwlD''. The genes ''cwlJ'' and ''sleB'' code for lytic enzymes which are active in the process of germination. In the work of Setlow et al, when ''cwlJ'' and ''sleB'' were knocked out together, germination frequency was reduced by 5 orders of magnitude.  When these genes are knocked out, we hope to yield a ''B. subtilis'' strain which produces spores incapable of germination.
-Our goal is to remove the germination capability, while keeping the necessary structural functions of the spores intact.
+Two methods are being employed to knock out germination: resistance cassette (RC) knockouts and clean deletions. Single RC knockouts were created, and have then been combined to create multiple knockouts.
-We have chosen genes involved in germination, and will knock them out to yield a ''B. subtilis'' strain which produces spores incapable of germination. This will be checked with germination assays.
+This will be checked with germination assays.
 Further, we will investigate the possibility of using a redundant toxin system to immediately kill off any spores which somehow germinate.