Team:LMU-Munich/Spore Coat Proteins/mainresult

From 2012.igem.org

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===GFP as a Proof of Principle===
===GFP as a Proof of Principle===
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<p align="justify">We obtained significant differences between wild type spores and all of our '''Sporo'''beads (see [http://2012.igem.org/Team:LMU-Munich/Data/gfp_spore data]). The intensity bar charts in Fig. 6 show the fluorescence intensity, while the 3D graphs illustrate the distribution of fluorescence intensity across the spore surface. This correlates with the localization of our fusion proteins in the crust. For image analysis we measured the fluorescence intensity of an area of 750 pixel per spore by using ImageJ and evaluated the results with the statistical software R.  
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<p align="justify">We designed the first GFP-'''Sporo'''bead to evaluate our system and compare different constructs.
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The following graph (Fig. 6) shows the results of microscopy and ImageJ analysis of the strongest construct integrated into wildtype W168 (B53) and the deletion strain B 49 (B70).</p>
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This short time lapse video shows the formation of a spore.</p>
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<p align="justify">To ensure that our '''Sporo'''beads show fluorescence on their edge and higher than in wildtype spores, we analysed the microscopy pictures. The following graph (Fig. 6) shows these microscopy pictures and analysis of the strongest of the five constructs integrated into wildtype strain (B53) and the deletion strain (B70).</p> 
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<font color="#000000"; size="2">Fig. 6: Result of fluorescence evaluation of the three strains W168, B53 and B70.</font>
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<font color="#000000"; size="2">Fig. 6: Result of fluorescence evaluation of the three strains W168, B53 and B70. The bar charts show the average fluorescence intensity over 100-200 spores. For image analysis we measured the fluorescence intensity of an area of 750 pixel per spore by using ImageJ and evaluated the results with the statistical software R. The 3D graphs illustrate in one spore the distribution of fluorescence intensity across the spore surface.</font>
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<p align="justify">As shown in Fig. 6, the wild type spore has hardly any fluorescence, whereas both''' Sporo'''beads with the integrated construct  pSB<sub>''Bs''</sub>1C-P<sub>''cotYZ''</sub>-''cotZ''<sub>-2aa</sub>-''gfp''-terminator give a distinct fluorescence signal around the edge of the spore. Furthermore, it demonstrates that strain B 70 has the highest fluorescence intensity. For more detailed information look at our [http://2012.igem.org/Team:LMU-Munich/Data/gfp_spore Data page]</p>
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<p align="justify">As shown in Fig. 6, the wild type spore has hardly any fluorescence, whereas both''' Sporo'''beads with the integrated construct  pSB<sub>''Bs''</sub>1C-P<sub>''cotYZ''</sub>-''cotZ''<sub>-2aa</sub>-''gfp''-terminator give a distinct fluorescence signal around the edge of the spore. Furthermore, it demonstrates that strain B 70 has the highest fluorescence intensity. For more detailed information have later look at our DATA page</p>
<p align="justify">In summary we successfully developed functional '''Sporo'''beads that are capable of displaying any protein of choice on the surface of modified ''B. subtilis'' endospores.</p>
<p align="justify">In summary we successfully developed functional '''Sporo'''beads that are capable of displaying any protein of choice on the surface of modified ''B. subtilis'' endospores.</p>
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[[File:NEXT.png|right|80px|link=Team:LMU-Munich/Application/Filtration_tour]]  [[File:BACK.png|left|80px|link=Team:LMU-Munich/Spore_Coat_ProteinsTour/Background]]
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[[File:NEXT.png|right|80px|link=Team:LMU-Munich/Application]]  [[File:BACK.png|left|80px|link=Team:LMU-Munich/Spore_Coat_ProteinsTour/Background]]
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Latest revision as of 16:48, 26 October 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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