Part:BBa_K2182003:Experience

Applications of BBa_K2182003

This part was introduced by Northeastern 2016 to characterize the Nox protein coding gene (BBa_K2182000) designed to scavenge oxygen in a microbial electrolysis chamber in order to increase efficiency and reduce the competitive advantage of E. coli over the facultative anaerobe Geobacter sulfurreducens.
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This dissolved oxygen assay was conducted to determine whether or not BBa_K2182003 was producing NOX sufficiently to impact dissolved oxygen levels. Over the course of the first 10 minutes the PBS containing BBa_K1282003 transformed Bl21 E. coli lost 35% of the initial dissolved oxygen whereas the PBS containing the control BL21 group saw only a 5% decrease. A CFU count was conducted in order to determine whether the observed difference in dissolved oxygen consumption was due only to a difference in number of cell counts between the control BL21 and the BBa_K1282003 transformed cells. Average CFU/ml were calculated using a dilution from the OD normalized liquid cultures. The mean CFU/ml of the liquid cultures are displayed in the table at the right and show that the BL21 culture had a higher CFU/ml than NOX did both as a raw mean and over the 95% confidence intervals of the two means. This demonstrates that BBa_K1282003 was causing a significant increase in oxygen consumption per cell compared to untransformed BL21 E. coli.

SDS-PAGE assays using His-purification were unsuccessful, possibly indicating low protein expression.

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