Edinburgh/Yoghurt/Status
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''Pseudomonas fluorescens'' culture collection: NCIMB 9046 | ''Pseudomonas fluorescens'' culture collection: NCIMB 9046 | ||
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| + | [[Edinburgh/Yoghurt| Introduction]] | [[Edinburgh/Yoghurt/Applications|Applications]] | [[Edinburgh/Yoghurt/Design|Design]] | [[Edinburgh/Yoghurt/Status|Status]] | [[Edinburgh/Yoghurt/References|References]] | ||
Latest revision as of 10:22, 9 August 2007
Edinburgh > Yoghurt
Introduction | Applications | Design | Status | References
https://static.igem.org/mediawiki/2007/f/f5/800px-Edinburgh_City_15_mod.JPG
Contents |
Status
Vanillin Genes
Sam5 & Sam8
- Saccharothrix espeanensis has arrived from DSMZ culture collection
- requires reviving and fcs and ech genes being PCR'd out
- Sam8 and Sam5 expression vector kindly donated by Dr Anton Linnenbrink from the Institute for Pharmazeutische Wissenschaften in Germany
- vector has been transformed into E. coli
- need to PCR out genes when primers arrive
COMT
- have grown up alfalfa seeds and genomic plant DNA has been extracted
- attempted to amplify up COMT via PCR, but have had no promising results so far
- RNA has been isolated from alfalfa
- have attempted to amplify up cDNA of COMT gene and are waiting on results
fcs & ech
- Pseudomonas fluorescens has arrived from NCIMB culture collection and has been revived
fcs
- have carried out several PCRs to amplify out the fcs gene from Pseudomonas fluorescenes
- PCRs did not result in one PCR product being produced, problem is most probably due to the P. fluorescens genome being of high GC content
- are attempting to purify out fcs gene from agrose gel of PCR products
ech
- the first set of primers ordered for ech did not work, as no PCR product was present after several PCR reactions
- have sent away for a second set of primers and are awaiting their arrival
vector
- we are going to modify a vector kindly donated by Norwich University, which can act as a shuttle vector between E. coli and a broad range of gram positive hosts, including Bacillis subtillis, Lactobacillus and Lactococcus
- firstly we will ligate MITs wintergreen scent generating device into the vector to characterise how the vector works in our chosen gram positive organisms
- secondly, when we have a working vanillin synthesis pathway, the vanillin biosynthesis pathway will be inserted into the vector and then transformed into Lactobacillus under the control of the arabinose promoter
- Lactobacillus may then be used to make yoghurt giving us self flavouring yoghurt, when the arabinose inducer is added
Enzyme Accession Numbers
Sam5 and Sam8 Accession: DQ357071
Saccharothrix espanaensis culture collection: DSM 44229
COMT Accession: M63853
fcs and ech Accession: DQ119298
Pseudomonas fluorescens culture collection: NCIMB 9046
Introduction | Applications | Design | Status | References
