Difference between revisions of "Part:BBa K1497013"

 
 
(4 intermediate revisions by 2 users not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1497013 short</partinfo>
 
<partinfo>BBa_K1497013 short</partinfo>
  
−
a
+
 
 +
<html>
 +
<div align="left">
 +
<table class="MsoTableGrid"
 +
style="border: 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: 306.7pt; height: 214.9pt;">
 +
The biobrick B0034-DFR is a composite consisting of the biobrick DFR (BBa_K1497013) and RBS (Ellowitz 1999, <a href="/Part:BBa_K1497023">BBa_B0034</a>).
 +
B0034 is a strong ribosome binding site with a biology efficiency of 1.0.<br>
 +
The dihydroflavonol 4-reductase (DFR; EC 1.1.1.219) from the plant <i> Dianthus caryophyllus </i> is an enzyme, catalyzing the reversible conversion of dihydroflavonols e.g. dihydro-kaempferol or dihydroquercetin into their corresponding leucoanthocyanidin. This reaction is NADPH-dependent (Liew et al. 1998) but its counter reaction can also occur with NAD<sup>+</sup> instead of NADP<sup>+</sup> (Queen Mary University of London 2014). DFR contains 353 amino acids and has a molecular weight of approximately 39.4 kDa. <br> <br>
 +
The iGEM Team TU Darmstadt 2014 used the B0034-DFR and verified the function of the B0034-DFR in their pelargonidin operon <a href="/Part:BBa_K1497023">(K1497023)</a>.
 +
</td>
 +
<td
 +
style="padding: 0cm 5.3pt; vertical-align: top; width: 136.7pt; height: 114.9pt;">
 +
      <img
 +
style="width: 500px; height: 150px;" alt=""
 +
src="https://static.igem.org/mediawiki/parts/a/ac/DFR_Wiki_reaction.png"></p>
 +
      <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 1</b></span></a><span lang="EN-US">
 +
Reaction of the DFR. One Dihydroflavonol reacts with NADPH to form a Leucoanthocyanidin. The reverse reaction works with NAD<sup>+</sup> or NADP<sup>+</sup>. </span></p>
 +
      </td>
 +
    </tr>
 +
<tbody>
 +
</table>
 +
</div>
 +
</html>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 17: Line 43:
 
<partinfo>BBa_K1497013 parameters</partinfo>
 
<partinfo>BBa_K1497013 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
====References====
 +
 +
1. Petit P, Granier T, d’Estaintot BL, et al. (2007) Crystal structure of grape dihydroflavonol 4-reductase, a key enzyme in flavonoid biosynthesis. Journal of molecular biology 368:1345–57. doi: 10.1016/j.jmb.2007.02.088
 +
 +
2. Gollop R, Even S, Colova-tsolova V, et al. (2002) Expression of the grape dihydroflavonol reductase gene and analysis of its promoter region 1. 53:1397–1409.
 +
 +
3. Liew C, Loh C, Goh C, Lim S (1998) The isolation , molecular characterization and expression of dihydroflavonol 4-reductase cDNA in the orchid , Bromheadia. 135:161–169.

Latest revision as of 01:03, 18 October 2014

B0034-DFR


The biobrick B0034-DFR is a composite consisting of the biobrick DFR (BBa_K1497013) and RBS (Ellowitz 1999, BBa_B0034). B0034 is a strong ribosome binding site with a biology efficiency of 1.0.
The dihydroflavonol 4-reductase (DFR; EC 1.1.1.219) from the plant Dianthus caryophyllus is an enzyme, catalyzing the reversible conversion of dihydroflavonols e.g. dihydro-kaempferol or dihydroquercetin into their corresponding leucoanthocyanidin. This reaction is NADPH-dependent (Liew et al. 1998) but its counter reaction can also occur with NAD+ instead of NADP+ (Queen Mary University of London 2014). DFR contains 353 amino acids and has a molecular weight of approximately 39.4 kDa.

The iGEM Team TU Darmstadt 2014 used the B0034-DFR and verified the function of the B0034-DFR in their pelargonidin operon (K1497023).

Figure 1 Reaction of the DFR. One Dihydroflavonol reacts with NADPH to form a Leucoanthocyanidin. The reverse reaction works with NAD+ or NADP+.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 348
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 100
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 85


References

1. Petit P, Granier T, d’Estaintot BL, et al. (2007) Crystal structure of grape dihydroflavonol 4-reductase, a key enzyme in flavonoid biosynthesis. Journal of molecular biology 368:1345–57. doi: 10.1016/j.jmb.2007.02.088

2. Gollop R, Even S, Colova-tsolova V, et al. (2002) Expression of the grape dihydroflavonol reductase gene and analysis of its promoter region 1. 53:1397–1409.

3. Liew C, Loh C, Goh C, Lim S (1998) The isolation , molecular characterization and expression of dihydroflavonol 4-reductase cDNA in the orchid , Bromheadia. 135:161–169.