Team:LMU-Munich/Spore Coat Proteins/mainresult

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Revision as of 15:17, 26 October 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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LMU Glow Spore2 cutII.jpg

GFP as a Proof of Principle

We designed the first Sporobead to express GFP to evaluate our system and compare different constructs. We had to ensure that production is both high and takes place on the edge of spores. The following graph (Fig. 6) shows the microscopy pictures and analysis of the strongest of the five constructs integrated into wildtype W168 (B53) and the deletion strain B 49 (B70).

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.

As shown in Fig. 6, the wild type spore has hardly any fluorescence, whereas both Sporobeads with the integrated construct pSB_Bs1C-P_cotYZ-cotZ_-2aa-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 Data page

In summary we successfully developed functional Sporobeads that are capable of displaying any protein of choice on the surface of modified B. subtilis endospores.

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

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 ===GFP as a Proof of Principle===
-<p align="justify">We obtained significant differences between wild type spores and all of our '''Sporo'''beads (see [https://2012.igem.org/Team:LMU-Munich/Data/gfp_spore data]). The following graph (Fig. 6) shows the microscopy pictures and analysis of the strongest of the five constructs integrated into wildtype W168 (B53) and the deletion strain B 49 (B70).</p>
+<p align="justify">We designed the first '''Sporo'''bead to express GFP to evaluate our system and compare different [[Team:LMU-Munich/Spore_Coat_Proteins/cloning|constructs]]. We had to ensure that production is both high and takes place on the edge of spores. The following graph (Fig. 6) shows the microscopy pictures and analysis of the strongest of the five constructs integrated into wildtype W168 (B53) and the deletion strain B 49 (B70).</p>
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