Difference between revisions of "Part:BBa K5120000"
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===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K5120000 SequenceAndFeatures</partinfo> | <partinfo>BBa_K5120000 SequenceAndFeatures</partinfo> | ||
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| + | ===Proof of Function=== | ||
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| + | [[File:projectdescription1.png|400px|thumb|left|'''Figure 2:''' Figure 2. HPLC chromatogram showing the detection of puerarin, daidzin, genistin, iso-vitexin, daidzein and genistein in transformed <i>N. benthamiana</i> samples ]] | ||
| + | [[File:projectdescription1.png|400px|thumb|left|'''Figure 3:''' Figure 3, concentration levels of three compounds—daidzein, genistein, and genistin—in Nicotiana benthamiana]] | ||
| + | |||
| + | <p>This part was used in a composite part along with other key enzymes in the isoflavonoid biosynthetic pathway and was agroinfiltrated into <i>N. benthamiana</i> using Agrobacterium tumefaciens. </p> | ||
| + | |||
| + | <p> | ||
| + | After transformation, the modified plants were tested for isoflavonoid production using High-Performance Liquid Chromatography (HPLC). The chromatogram shows the amounts of each target isoflavonoid: puerarin, daidzein, and genistein with the first peak, observed at around 16.0 minutes, representing puerarin, followed by a peak at approximately 17.0 minutes, which corresponding to daidzin. Further along, a peak at 22.0 minutes is attributed to genistin. Traces of all three compounds were detected in <i>N. benthamiana</i>, a plant that does not naturally produce any of these because it lacks the enzymes needed to do so. | ||
| + | |||
| + | This shows that the sequence for CHS did function as intended because if it hadn't then the pathway wouldn't have progressed further and produced these isoflavonoids. | ||
| + | </p> | ||
| + | |||
===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K5120000 parameters</partinfo> | <partinfo>BBa_K5120000 parameters</partinfo> | ||
Revision as of 03:32, 1 October 2024
Chalcone Synthase (CHS) in Pueraria Mirifica Var. Lobata
Usage and Biology
Chalcone Synthase (CHS) is an enzyme that catalyzes the first step in the production of flavonoids, converting p-coumaroyl-CoA and malonyl-CoA into naringenin chalcone or isoliquiritigenin, which is the precursor for various flavonoid compounds, including flavanones, flavonols, and anthocyanins.
It is constructed for use in plants, specifically Nicotiana benthamiana, to be included in a composite part that allows for the expression of CHS in the biosynthesis of isoflavonoids.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Proof of Function
This part was used in a composite part along with other key enzymes in the isoflavonoid biosynthetic pathway and was agroinfiltrated into N. benthamiana using Agrobacterium tumefaciens.
After transformation, the modified plants were tested for isoflavonoid production using High-Performance Liquid Chromatography (HPLC). The chromatogram shows the amounts of each target isoflavonoid: puerarin, daidzein, and genistein with the first peak, observed at around 16.0 minutes, representing puerarin, followed by a peak at approximately 17.0 minutes, which corresponding to daidzin. Further along, a peak at 22.0 minutes is attributed to genistin. Traces of all three compounds were detected in N. benthamiana, a plant that does not naturally produce any of these because it lacks the enzymes needed to do so. This shows that the sequence for CHS did function as intended because if it hadn't then the pathway wouldn't have progressed further and produced these isoflavonoids.