Part:BBa_K917005:Experience

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Applications of BBa_K917005

Expression of this biobrick inhibits growth of cells in the presence of nitro drugs such as metronidazole. We have rigorously tested this biobrick's potential as a counter-selectable marker in its pre-biobrick form (in Blue script vector with lac promoter). We believe that this is a good representation of the biobrick's activity since the specific nitroreductase activity of this biobrick is very similar to the specific nitroreductase activity in the blue script vector:

Comparison of the specific activity of biobricked nitroreductase gene and control under induced and uninduced conditions (+ and - IPTG respectively). Error bars are standard error of the mean.

Comparison of the specific activity of 3 nitroreductase genes into different vectors with different promoters and control. Error bars are standard error of the mean.

Although growth of cells containing nitroreductase is inhibited in plates, it is not absolute and therefore unsuitable for couter-selection.

Nitroreductase containing cells's growth (right) is markedly inhibited in comparison to control cells (left) at 90 ug/ml metronidazole. However, this inhibition is not absolute so cannot be used for counterselection.

Aerobic liquid culture results are more promising with profound growth inhibition of nitroteductase-containing cells while control cells are growing.

Liquid aerobic cultures of BS-nitroreductase and BS-control in the presence and absence of metronidazole. At 150 ug/ml metronidazole, cells which contain nitroreductase show almost no growth.The measurements were made in triplicate. Error bars are standard error of the mean. Inital OD600 (just after inoculation) was subtracted.

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