Team:TU-Delft/part3

Introduction

By combining the olfactory receptor and the FUS1pr-EGFP reporter, a complete yeast olfactory system is obtained. If the corresponding ligand binds to the receptor the FAR1 promoter is turned on and the EGFP is expressed. This EGFP signal can be read out by a fluorescence meter. If the olfactory system will be implemented as a diagnostics tool in developing countries, the EGFP reporter should be changed by a visible reporter.
Besides the induction of the FUS1 promoter the cells also go in growth arrest mediated by the FAR1 promoter. However it is undesirable that the cells stop growing once they respond to a ligand. Therefor it is needed to knock out the FAR1 promoter.

Parts

The receptor constructs and the reporter constructs are combined to have one complete olfactory system. The following biobricks are created:

BBa_K775005
BBa_K775006
BBa_K775007
BBa_K775008

Results

Knock out FAR1

Setup

Outcome

Fluorometer experiment

Setup

With yeast strains transformed with the GPR109A receptor and output BBa_K775005 and the R17-ODR10 receptor and output BBa_K775008 a fluorometer experiment was performed. After addition of the ligands OD600 and fluorescence were measured in time.

Outcome

Conclusions

Recommendations