From 2007.igem.org

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Revision as of 09:21, 11 October 2007

In order to test the feasibility of gene expression in Lactobacillus and the possibility of making self flavouring yoghurt, we required a proof of concept.

As both the pigment and flavour pathways are rather complex and will require modification and optimisation before their expression in yoghurt we are going to use the much simpler RFP gene as a proof of concept.

pTG262-PLac-RFP construct

We have inserted the lactose induced RFP gene into the pTG262 vector

• this vector was then transformed into E. coli
• growth on IPTG-Xgal media resulted in RFP expression and production of red colonies

Further experimentation

• transform pTG262-Plac-RFP vector into Lactobacillus bulgaricus and Steptococcus thermophilus
• plate colonies onto IPTG-Xgal media and see if RFP is expressed
• introduce LAB with pTG262-Plac-RFP vector into yoghurt started culture & make yoghurt
• see if yoghurt produced turns red

Presently we are transforming the LAB with the pTG262-Plac-RFP vector

pTG262-Pbad-RFP construct