Difference between revisions of "Part:BBa K4012029"
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[https://parts.igem.org/Part:BBa_K4012029] A: The results of colony PCR of upstream transformant; B: The results of colony PCR of downstream transformant; C: Construction of catechin metabolic pathway; D：Construction of naringenin metabolic pathway]] | [https://parts.igem.org/Part:BBa_K4012029] A: The results of colony PCR of upstream transformant; B: The results of colony PCR of downstream transformant; C: Construction of catechin metabolic pathway; D：Construction of naringenin metabolic pathway]] | ||
Assembly of synthesis pathway of naringenin, shown by Fig.1. | Assembly of synthesis pathway of naringenin, shown by Fig.1. | ||
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===Fermentation results by HPLC test=== | ===Fermentation results by HPLC test=== | ||
During the HPLC testing, we purchased final and intermediate metabolism products standard, naringenin, taxifolin, eriodictyol and catechin, and used them in the subsequent HPLC tests. | During the HPLC testing, we purchased final and intermediate metabolism products standard, naringenin, taxifolin, eriodictyol and catechin, and used them in the subsequent HPLC tests. | ||
Revision as of 14:08, 21 October 2021
RtTAL-Pc4CL-GbCHS-MsCHI level2
It is a composite part of naringenin metabolic pathway. pRET2-RtTAL-tSSA1-pPGK1-Pc4CL-tCYC1-pTEF1-GbCHS-tADH1-pPOP6-MsCHI-tTDH1
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 195
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 714
Illegal BglII site found at 2758
Illegal BglII site found at 4201
Illegal BglII site found at 6848
Illegal BglII site found at 9038
Illegal XhoI site found at 2844
Illegal XhoI site found at 8072
Illegal XhoI site found at 9746 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 50
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3
Illegal BsaI site found at 2830
Illegal BsaI site found at 3099
Illegal BsaI site found at 6137
Illegal BsaI site found at 8058
Illegal BsaI site found at 8327
Illegal BsaI.rc site found at 724
Illegal BsaI.rc site found at 3080
Illegal BsaI.rc site found at 4211
Illegal BsaI.rc site found at 6858
Illegal BsaI.rc site found at 8308
Illegal BsaI.rc site found at 9048
Illegal SapI site found at 7762
Illegal SapI.rc site found at 7711
Level2 plasmid assembly
Figure 1: [1] A: The results of colony PCR of upstream transformant; B: The results of colony PCR of downstream transformant; C: Construction of catechin metabolic pathway; D：Construction of naringenin metabolic pathway
Assembly of synthesis pathway of naringenin, shown by Fig.1.
Fermentation results by HPLC test
During the HPLC testing, we purchased final and intermediate metabolism products standard, naringenin, taxifolin, eriodictyol and catechin, and used them in the subsequent HPLC tests.
Figure 2: [2] Fig.2 The results of HPLC of naringenin, taxifolin, eriodictyol, catechin standard and sample from fermentation medium
As shown in Fig.2, since we could not buy leucoanthocyanidins (LCD), we only did HPLC of the four intermediates: Catechin, Eriodictyol, Taxifolin and Naringenin. Catechin and other intermediate metabolites were confirmed by comparing the samples with the standard.