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Team:KULeuven/Lab/Vanillin Production
From 2009.igem.org
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==Steps== | ==Steps== | ||
*we will be working in three different stages | *we will be working in three different stages | ||
| - | 1.Testing the transformation of tyrosin to ferulic acid. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC | + | 1.Testing the transformation of tyrosin to ferulic acid. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC |
| - | 2.Testing the transformation of ferulic acid to vanillin. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC | + | 2.Testing the transformation of ferulic acid to vanillin. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC |
| - | 3.If both are tested and ok, they can be combined into one plasmid and again tested. | + | 3.If both are tested and ok, they can be combined into one plasmid and again tested. |
| - | 4.Then, the locks need to be added. | + | 4.Then, the locks need to be added. |
*testing the enzymes: by detecting the endproducts through gaschromatography. | *testing the enzymes: by detecting the endproducts through gaschromatography. | ||
*One problem might be that the tyrosin of E.Coli might be depleted to fast | *One problem might be that the tyrosin of E.Coli might be depleted to fast | ||
Revision as of 13:42, 24 July 2009
Contents |
VANILLIN SYNTHESIS
Goal
Making vanillin from tyrosine in a five-step pathway.
required
- biobricks:
SAM8: BBa_ I742142: coding sequence without RBS SAM5: no number: RBS site + PstI restriction site removed COMT: BBa_I742107: coding sequence without RBS FCS: BBa_I742115: RBS + fcs in pSB1A2 ECH: BBa_I742113: RBS + ech in pSB1A2
Where from
- 1. COMT is available in 2009 spring distribution plate 1 well 19D
DNA is compatible
- 2. SAM8, SAM5, COMT, FCS, ECH will be sent to us from the French Lab from the university Edinburgh
Steps
- we will be working in three different stages
1.Testing the transformation of tyrosin to ferulic acid. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC 2.Testing the transformation of ferulic acid to vanillin. The enzymes are put under a constitutive promoter and transcribed. Ferulic acid concentrations are then measured with GC 3.If both are tested and ok, they can be combined into one plasmid and again tested. 4.Then, the locks need to be added.
- testing the enzymes: by detecting the endproducts through gaschromatography.
- One problem might be that the tyrosin of E.Coli might be depleted to fast
