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<p>In combing Bacillus subtilis powerful secretion capacity with Peptidosomes as a new platform for functional co-cultivation we aim to produce multi protein complexes. Various strains - each secreting distinct proteins of interest - can be cultivated in one reaction hub while being physically separated. In this part of EncaBcillus we study extracelluar protein interaction mediated by the SpyTag/SpyCatcher system. This set-up bears the potential for an effective production of customizable biomaterials or enzyme complexes.</p> | <p>In combing Bacillus subtilis powerful secretion capacity with Peptidosomes as a new platform for functional co-cultivation we aim to produce multi protein complexes. Various strains - each secreting distinct proteins of interest - can be cultivated in one reaction hub while being physically separated. In this part of EncaBcillus we study extracelluar protein interaction mediated by the SpyTag/SpyCatcher system. This set-up bears the potential for an effective production of customizable biomaterials or enzyme complexes.</p> | ||
<h2>Achievments</h2> | <h2>Achievments</h2> | ||
+ | <div style="display: flex; align-items: flex-start; flex-wrap: wrap;"> | ||
+ | <div class="makeresponsive" style="width:50%;"> | ||
+ | <figure class="makeresponsive" style="padding-left:10%; padding-right:10%;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/0/08/T--TU_Dresden--secretion--demonstrate1.png" | ||
+ | alt="Figure 1: Endpoint measurement of the fluorescence from supernatants." | ||
+ | class="makeresponsive zoom"> | ||
+ | <figcaption><b>Figure 1: Endpoint measurement of the fluorescence from supernatants. </b> | ||
+ | Expression of the multi copy mCherry constructs (purple) was induced with 1% Xylose and the supernatants were harvested after 16 h of incubation. Wild type supernatant is shown as a control (pink). Excitation wavelength was set to 585 nm and emission was recorded at 615 nm. The fluorecense was normalized over the optical density of the cell culture at 600 nm (OD600). Graph shows mean values and standard deviations of at least two biological and three technical replicates. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | <div class="makeresponsive" style="width:50%;"> | ||
+ | <figure class="makeresponsive floatleft" style="padding-left:10%; padding-right:10%;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/b/b7/T--TU_Dresden--secretion--results5.png" | ||
+ | alt="Figure 5: Supernatants of <i>B. subtilis</i> cultures." | ||
+ | class="makeresponsive zoom"> | ||
+ | <figcaption><b>Figure 5: Supernatants of <i>B. subtilis </i>cultures.</b> | ||
+ | Wild-type supernatant (left) and a mCherry-mini. SpyCatcher secreting strain (right). The expression of the multi-copy mCherry was induced with 1% Xylose and the supernatant was harvested after 16 h of incubation. | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </div> | ||
+ | </div> | ||
</div> | </div> | ||
Revision as of 18:46, 28 October 2017