Team:INSA-Lyon/Results

From 2014.igem.org

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<h6> Transmission Electron Microscopy
<h6> Transmission Electron Microscopy
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<img src="https://static.igem.org/mediawiki/2014/4/4f/Insa_labo.jpg" alt="les filles au labo"  
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<p>For the Transmission Electron Microscopy, all the strains were cultivated in conditions that allow the formation of the curli : 48h (for a 50 mL culture),  28⁰C temperature and low agitation. The ammonium molybdate was used as a negative colorant (Microscope: MET PHILIPS CM120). </p>
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<img src="https://static.igem.org/mediawiki/2014/8/8a/Figure5_MET.png"  
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<p><b>Figure 5: Engineered bacteria curli structure observation using Transmission Electron Microscopy</b></p>
<p><b>Figure 5: Engineered bacteria curli structure observation using Transmission Electron Microscopy</b></p>
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<p>For the Transmission Electron Microscopy, all the strains were cultivated in conditions that allow the formation of the curli : 48h (for a 50 mL culture),  28⁰C temperature and low agitation. The ammonium molybdate was used as a negative colorant (Microscope: MET PHILIPS CM120). </p>
 
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<p>The bacteria cultures we used are a csgA- strain as negative control, the strain that naturally does the curli and the two engineered csgA constructions, the His-Tag and His2-Tag. </p>
 
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<p>The images show that there is no significant difference between our positive control and our constructions.  Thus, we can conclude that our parts insertions don’t affect the structure of the amyloid fibers and the configuration of the curli formation. </p>
 
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<p>The bacteria cultures we used are a <i>csgA</i>-knockout strain as negative control, and the three engineered csgA constructions, the WT, the His1-Tag and His2-Tag. </p>
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<p>The images show that there is <b>no significant difference between our positive control and our constructions</b>.  Thus, we can conclude that <b>our parts insertions don’t affect the structure of the amyloid fibers and the configuration of the curli formation</b>. </p>
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<p><br/><br/>We can conclude that <b>expression of CsgA derivatives</b> from the p70 <i>csg</i> promoter carried by the psb1c3 plasmid leads to <b>functional CsgA</b>, and allows <i>E. coli</i> to <b>stick and form biofilm</b>. Moreover, our results show that the <b>addition of one or two His-Tag on C-term of CsgA doesn’t disturb the normal properties of curli</b> (sturdiness, adhesion and folding of CsgA).</p></div></li>
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<p><br/><br/>As a conclusion, <b>the expression of CsgA derivatives</b> from the p70 <i>csg</i> promoter carried by the psb1c3 plasmid leads to <b>functional CsgA</b>, and allows <i>E. coli</i> to <b>stick and form biofilm</b>. Moreover, our results show that the <b>addition of one or two His-Tag on C-term of CsgA doesn’t disturb the normal properties of curli</b> (sturdiness, adhesion and folding of CsgA).</p></div></li>
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Revision as of 19:56, 17 October 2014

Curly'on - IGEM 2014 INSA-LYON

  • Curli characterization


  • Nickel chelation


  • Survival after UV and high temperature exposure


  • Promoter optimization and characterization