Team:Carnegie Mellon/Project/Results

KillerRed phototoxicity

Our photobleaching experiments showed the phototoxicity of KillerRed in pSB1A3:

Figure 1: Phototoxicity of KillerRed and mRFP1 (<partinfo>BBa_E1010</partinfo>) on E. coli XL10 by irradiating with a HBO100 lamp (includes 375 nm LP filter) for 5 hours.

KillerRed's phototoxic effect on E. coli XL10 is shown in Figure 1.

Triplicate cell viabilities are divided by the normalized fluorescence values to account for bleaching effects and the fact that KillerRed has a lower expression level than mRFP1. RFP 114% ± 20% (viable cells)/(Normalized RFU)
KillerRed: 53% ± 17% (viable cells)/(Normalized RFU)

KillerRed photobleaching curve

Figure 1: Photobleaching curve of KillerRed with a HBO100 mercury-arc lamp

XL10 Ultracompetent cells were transformed with KillerRed (BBa_K1184000) cloned with <partinfo>BBa_B0034</partinfo> as the RBS and <partinfo>BBa_R0010</partinfo> as the wild-type lac promoter and induced overnight with IPTG.The overnight was bleached for 180 minutes with HBO100 (100W Mercury-arc lamp). Fluorescence data was taken using a Tecan Safire II with the parameters shown in Table 1. Fluorescence Data is shown in Table 2.

To read our conclusions and future directions, visit our Abstract and Future Directions pages.