Team:NEYCFLS China/Parts
- Central Metabolism Pathway/Aromatic Pathway
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The E.Coli used in for this experiment is strain W3110/ATCC 23725. The central metabolism pathway and the common aromatic pathway, where glucose is condensed to PEP, DAPH and to CHA uses the following primers: CTG CAG GTG ATG ATT ATC AGC CAG CAG and GGA TCC CAA TGC TTC GTT TCG TAT CAC. To amplify the effects of the pathway, genomic DNA of E. coli W3110 was enhanced using the primers AGA TCT ATG CAA AAA GAC GCG CTG, AGA TCT TTA AGC CAC GCG AGC CGT and GGA TCC ATG CAA ACA CAA AAA CCG ACT, GGA TCC TTA CCC TCG TGC CGC CAG.
- Central Metabolism Pathway/Aromatic Pathway
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In E.Coli, there are enzymes that convert chorismate into tryptophan, phenylalanine, and tyrosine. For us, we also need to block the genes for pheA and tyrA to ensure that we only acquire Tryptophan as a final product.
Primers for pheA: CTC CCA AAT CGG GGG GTT TTT TAT TGA TAA CAA AAA GGC AAC ACT ATG, GAT GAT TCA CAT CAT CCG GCA CCT TTT CAT CAG GTT GGA TCA ACA GGA
Primers for tyrA: GAT GAT GTG AAT CAT CCG GCA CTG GAT TAT TAC TGG CGA TTG TCA TTC, GGA TCT GAA CGG GCA GCT GAC GGC TCG CGT GGC TTA AGA GGT TTA TTA
- Chorismate to Indole
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E.Coli has all the main ingredients for converting chorismate to tryptophan.
Primers for trpBA: AGA TCT ATG CAA AAA GAC GCG CTG, AGA TCT TTA AGC CAC GCG AGC CGT
Primers for trpA: AAC CGG GGG AGG CAT TTT GCT TCC CCC GCT AAC AAT GGC GAC ATA TTA, ATG CGC CAC GTC TTA TCA GGC CTA CAA AAT CAA TCG CTT TTC AGC AAC
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Please note that in this file, the primers were grouped in small groups of 3 to prevent problems with the html code. This does not represent the way the primers are found in bacteria.
This page introduces all of the strains and genes that we (theoretically) worked on during the course of iGEM 2021. Because we were not able to get in a laboratory, we could only get the genetic information and strain names from articles we bought from Google Scholar. The photo below is the complete diagram for E.Coli converting glucose into indole in 3 major steps.
