Difference between revisions of "Part:BBa K862000"

Revision as of 16:02, 24 June 2012

precA-LacZ-doubleTerminator

This is a part for the precise quantification of UV-radiation or radioactive radiation in E. coli (recA+) strains, i.e. BL21(DE3). It consits of a recA Promoter (part BBa_J22106) fused to a LacZ reporter cloned in front of a double terminator (part BBa_K173004).

Characterisation

This part was characterized during the 2012 HS competition by the Heidelberg LSL team

Fig. 1: ONPG assay of Bl21(DE3) irradiated for different times. Both constructs show a strong correlation between the UV-irradiation time and the LacZ activity (production of o-nitrophenol).

Fig. 2: X-gal assay of Bl21(DE3) transformed with precA/precB/psulA_LacZ parts and irradiated for different times. All constructs show a strong positive correlation between UV induction time and coloring of the wells. PrecA-LacZ gives the lowest reporter background expression whereas psulA-LacZ gives the highest overall coloring of the samples.

Fig. 3: Test of precA-GFP reporter construct by fluorescence microscopy. E.coli transformed with our precA-GFP reporter were either UV-irradiated for 30 min or left uninduced. measurement of GFP expression show a 10-fold (!) increase in GFP expression due to UV-irradiation compared to the control.

Sequence and Features