Part:BBa_J100307:Experience

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Applications of BBa_J100307

J100307 was used to build part BBa_J100308, a variant of repClone Red with wt TetR promoter. We transformed E. coli with plasmids containing J100308 or J100306 (wt repClone Red with wt tetR promoter) and measured the induction caused by the addition of 200 ng/mL anhydrous tetracycline (aTc) to determine the incubation time needed for maximum induction. We found that aTc is most effective after fourteen to sixteen hours of growth, after which the induction is lost.

We performed a dose-response test on E. coli containing part J100306 to determine the ideal concentration of aTc for induction of RFP. We found that 400 ng/mL aTc gives the best compromise of fold induction and allowing for sufficient cell growth. Since J100308 and J100306 responded similarly in the time-response test, we expect that 400 ng/mL aTc would also be ideal for J100308 and other repClone constructs built from the variant J100307. Induction above 400 ng/mL looks higher because fewer cells grew and therefore the denominator was smaller when calculating the ratio.

Results of Dose Response Assay:

Results of Time Assay:

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