Difference between revisions of "Part:BBa K1497013"

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.

@@ Line 14: / Line 14: @@
 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<sub>+</sub> instead of NADP<sub>+</sub> (Queen Mary University of London 2014). DFR contains 353 amino acids and has a molecular weight of approximately 39.4 kDa. <br> <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>.
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