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2019 iGEM team Linkoping Sweden

2019 iGEM team Linkoping Sweden validated this part.

Figure 1. To the left This biobrick after 48 hours in 37 degrees Celsius (both pictures). To the left is cultured E. coli BL21 cells with this biobrick in white light. The culture tubes had a constant supply of oxygen (cotton plugs) which is important for the chromophore/fluorophore of eforRed to develop. To the top right A culture of BL21 (DE3) Gold which has been incubated for 48 hours in 37 degrees Celsius has been centrifuged at 12 000 g for 10 min. The result is a pellet of eforRed expressing bacteria with a burgundy color. The same pellet in the bottom right was put on a UV table (302 nm) which resulted in a pink glowing pellet.

To the right is colonies in the same host as before illuminated in 302 nm UV-light, showing eforReds ability to also emit strong fluorescence.

Figure 2. To the left is a spectrophotometric experiment where varying levels of oxygen supply were tested. The holes were made in the plastic cover of a 96-well plate and run for 16 hours in 37 degrees Celsius, this results shows the oxygen dependency of eforReds chromophore/fluorophore development. To the right is this biobrick in a 96-well plate, the culture has been in the well O.N in 37 degrees Celsius with varying levels of oxygen supply. The plate to the right is different expression systems used to test a strong green/yellow fluorescent protein. These results show eforReds ability to fluorescence, making it a fluorescent as well as a chromoprotein. Both plates were illuminated in 302 nm UV-light.

Figure 3. SDS-page of the sonicated BL21(de3) lysate with eforRed. Biorads "Precision Plus Protein Dual Color Standards" was used as the protein ladder. The visible band on the gel lies between 25 and 37 kD which correspond to the molecular weight of eforRed.

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