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Part:BBa_K917005:Experience

Designed by: Elitsa Peeva   Group: iGEM12_Edinburgh   (2012-09-20)
Revision as of 16:10, 25 September 2012 by Pandeli (Talk | contribs) (→‎Applications of BBa_K917005)

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

Nitroreductase_plates.jpg

Nitroreductase containing cells's growth (right) is markedly inhibited in comparison to control cells 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.

Nitroreductase_aerobic_culture.jpg

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. Error bars are standard error of the mean. Inital OD600 (just after inoculation) was subtracted.

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