- Cell sensor constructs which were based on CooA under T7 promoter in pTriEx vector transformed into E.coli BL21 DE3 host have failed to function.
- Alternative sensor construct with pCooM promoter/RFP signal and CooA under pLac promoter in pSB1C3 vector transformed into E.coli Top10 host worked successfully ! E.coli cells transformed with this construct produces very noticable RFP signal -even by naked eye- when exposed to carbon monoxide.
- 16 biobricks are constructed and submitted to the registry of standart parts. 12 of 16 parts are sent to IGEM HQ as DNA samples.
- CooA overexpression and purification procedures were optimized.
- CooA binds to response elements in the absence and presence of CO as indicated by EMSA studies and basal level GFP/RFP expression. On the other hand,
there is a significant increase in expression levels when CO activates CooA.
Human Practice
- We hope to provide a technical guideline for other future gas sensing cell sensor projects with our E-CO sensor with enhanced dynamic range project.