Difference between revisions of "Part:BBa K1497020:Experience"

 
(→‎Applications of BBa_K1497020)
 
Line 5: Line 5:
  
 
===Applications of BBa_K1497020===
 
===Applications of BBa_K1497020===
 +
The Biobrick <html><a href="/Part:BBa_K1497019">BBa_K1497019</a></html> produces in <i>E. coli</i> B and K strains the FdeR Protein. The iGEM Team TU Darmstadt 2014 measured the fluorescense of GFP and mKate after the incubation with diffrent conentrations of naringenin. The results are shown in Figure 3. 
 +
 +
<html>
 +
<div align="center">
 +
<table class="MsoTableGrid"
 +
style="border: medium none ; border-collapse: collapse; text-align: left; margin-left: auto; margin-right: auto;"
 +
border="0" cellpadding="0" cellspacing="0">
 +
  <tbody>
 +
    <tr style="height: 214.9pt;">
 +
      <td
 +
style="padding: 0cm 5.4pt; vertical-align: top; width: 236.7pt; height: 214.9pt;">
 +
      <img
 +
style="width: 800px; height: 342px;" alt=""
 +
src="https://static.igem.org/mediawiki/2014/3/32/Narinmrfpgfp.png"></p>
 +
      <br>
 +
      <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 3</b></span></a><span lang="EN-US">
 +
<b>Left:</b> Characterization of  <a href="/Part:BBa_K1497020">BBa_K1497020</a>. GFP fluorescence depends on the concentration of naringenin. We measured the GFP fluorescence after 16 h incubation with different concentrations of naringenin. By setting higher concentrations of naringenin, we gained higher fluorescence of GFP as well. <b>Right:</b> Characterization of <a href="/Part:BBa_K1497021">BBa_K1497021</a>. mKate (<a href="/Part:BBa_K1055000">BBa_K1055000</a>) fluorescence depends on the concentration of naringenin. We measured the mKate (<a href="/Part:BBa_K1055000">BBa_K1055000</a>) fluorescence after 16 h incubation with different concentrations of Naringenin. By setting higher concentrations of naringenin, we gained higher fluorescence of mKate as well.</span></p>
 +
      </td>
 +
    </tr>
 +
<tbody>
 +
</table>
 +
</div>
 +
</html>
 +
<html>
 +
 +
 +
 +
<br><br>
 +
</html>
 +
 +
 +
====In vivo characterisation of the naringenin biosynthesis operon (<html><a href="/Part:BBa_K1497007">BBa_K1497007</a></html>)====
 +
 +
<html>
 +
<div align="right">
 +
<table class="MsoTableGrid"
 +
style="border:1 medium none ; border-collapse: collapse; text-align: left;"
 +
border="0" cellpadding="0" cellspacing="0">
 +
  <tbody>
 +
    <tr style="height: 214.9pt;">
 +
   
 +
<td style="padding: 0cm 5.4pt; vertical-align: top; width: 506.7pt; height: 214.9pt;"><br>
 +
iGEM TU Darmstadt 2014 reconstitute the naringenin biosynthesis in <i>E. coli</i> by construction of a operon polycistronic gene cluster (<a href="/Part:BBa_K1497007">BBa_K1497007</a>) under control of the strong T7 promoter (<a href="/Part:BBa_K1497017">BBa_K1497017</a>). They used the naringenin biosensor with GFP response <a href="/Part:BBa_K1497020">K1497020</a> to characterize the naringenin biosynthesis operon in <i>E. coli</i> BL21(DE3). <br><br> The result are shown in figure 4. The GFP fluorescene is only in the cells with the T7 naringenin operon visible and detectable. The team determined for this operon a naringenin production yield of 3 µmol naringenin per liter.   
 +
 +
</td>
 +
 +
 +
 +
 +
<td
 +
style="padding: 0cm 5.4pt; vertical-align: top; width: 136.7pt; height: 114.9pt;">
 +
 +
      <img src="https://static.igem.org/mediawiki/2014/1/12/Naringenint7balken.png" align="right"
 +
  height="200" width="500" style="margin-right: 20px;"/></p>
 +
      <br><br>
 +
      <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 4</b></span></a><span lang="EN-US">
 +
<b>Left: </b>Cell pellets with and without T7-Naringenin operon from <i>E. coli</i> BL21(DE3)-pSB1C3-<i>fdeR-gfp</i>. By using ultraviolet light the pellet containing the naringenin operon shows a GFP fluorescence. <b>Right: </b>Measurement of the GFP fluorescence in the<i>E. coli</i> BL21(DE3)-pSB1C3-<i>fdeR-gfp</i> strain containing and not containing the T7-Naringenin operon.
 +
</span></p>
 +
      </td>
 +
 +
 +
 +
    </tr>
 +
<tbody>
 +
</table>
 +
</div>
 +
</html>
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 21:15, 17 October 2014


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1497020

The Biobrick BBa_K1497019 produces in E. coli B and K strains the FdeR Protein. The iGEM Team TU Darmstadt 2014 measured the fluorescense of GFP and mKate after the incubation with diffrent conentrations of naringenin. The results are shown in Figure 3.


Figure 3 Left: Characterization of BBa_K1497020. GFP fluorescence depends on the concentration of naringenin. We measured the GFP fluorescence after 16 h incubation with different concentrations of naringenin. By setting higher concentrations of naringenin, we gained higher fluorescence of GFP as well. Right: Characterization of BBa_K1497021. mKate (BBa_K1055000) fluorescence depends on the concentration of naringenin. We measured the mKate (BBa_K1055000) fluorescence after 16 h incubation with different concentrations of Naringenin. By setting higher concentrations of naringenin, we gained higher fluorescence of mKate as well.



In vivo characterisation of the naringenin biosynthesis operon (BBa_K1497007)


iGEM TU Darmstadt 2014 reconstitute the naringenin biosynthesis in E. coli by construction of a operon polycistronic gene cluster (BBa_K1497007) under control of the strong T7 promoter (BBa_K1497017). They used the naringenin biosensor with GFP response K1497020 to characterize the naringenin biosynthesis operon in E. coli BL21(DE3).

The result are shown in figure 4. The GFP fluorescene is only in the cells with the T7 naringenin operon visible and detectable. The team determined for this operon a naringenin production yield of 3 µmol naringenin per liter.



Figure 4 Left: Cell pellets with and without T7-Naringenin operon from E. coli BL21(DE3)-pSB1C3-fdeR-gfp. By using ultraviolet light the pellet containing the naringenin operon shows a GFP fluorescence. Right: Measurement of the GFP fluorescence in theE. coli BL21(DE3)-pSB1C3-fdeR-gfp strain containing and not containing the T7-Naringenin operon.

User Reviews

UNIQc45b14c39fcf8e03-partinfo-00000005-QINU UNIQc45b14c39fcf8e03-partinfo-00000006-QINU