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| | ===Design Notes=== | | ===Design Notes=== |
| − | To analyze the pelagonidin production operon (<html><a href="/Part:BBa_K1497015">K1497015</a></html>), we transformed it into <i>E.coli</i> Bl21(DE3). An overnight LB culture was used to inoculate an expression-culture. The expression of pelargonidin was performed according to Yan et al., (2007). After the induction with 1 mM Isopropyl-β-D-thiogalactopyranosid (IPTG) <i>E.coli</i> BL21 (DE3) cells were transferred into M9-media and fermented for 48h at 37°C in present of 0.1 mM naringenin.
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| − | src="https://static.igem.org/mediawiki/parts/d/d7/Pelletf%C3%A4rbungII.png"></p>
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| − | <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 2</b></span></a><span lang="EN-US">
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| − | <i>E.coli</i> BL21 (DE3) pellet containing the pelargonidin producing operon after the fermentation. According to Yan et al. (2007) a pelargonidin producing <i>E.coli</i> should be red after a pelargenidin production. The operon with the engineered anthocyanindin synthase produces more pelargonidin</span></p>
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| − | After the expression of pelagonidin producing operon with engineered ANS (<html><a href="/Part:BBa_K1497015">K1497015</a></html>) in present of 0.5 mM narigenin we performed an extraction of pelargonidin with methanol /dichloromethane from the pellet and supernatant and verified the pH-dependency of pelargonidin (Fig. 3).
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| − | <p class="MsoCaption" align="text-align:justify"><span lang="EN-US"><b>Figure 3</b></span></a><span lang="EN-US">
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| − | Extracted pelargonidin from <i>E.coli</i> BL21 (DE3) under day light. The color of pelargonidin depends on pH value and solvent. This indicates the present of pelargonadin. Left: Methanol extraction; right: Dichlormethane extraction.</span></p>
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