Part:BBa_K649201:Experience

Applications of BBa_K649201

We prepared a competent cell JM2.300 into which Pbad/araC-Cre(pSB1A2, BBa_I718008) had been constructed. Subsequently, the BioBrick was constructed into the cell. The strain was grown in a 3ml liquid culture, and 75μl of 2M arabinose was added to induce Cre expression. For the control experiments we used the same strain without arabinose induction and a JM2.300 strain which was induced with arabinose and had only our BioBrick. All the strains were cultured each for periods of 0.5, 1, 2, and 4 hours, and in each case the florescence levels were measured by flow cytometer and FLC.

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We measured the fluorescence levels to prove that recombination only occurs in presence of Cre recombinase. On the sample with the Pbad/araC-Cre construction, we found that recombination occurred when arabinose was added. We could also observe recombination occurred when arabinose was not added, which can be explained due to a leaking in the Pbad/araC promoter. Opposite to this result, when we measured the levels of the sample without the Pbad/araC-Cre construction, we found that the GFP levels were far lower than those of the sample with the Pbad/araC-Cre construction. This clearly proves that our lox constructions, both in BBa_K649201 and BBa_K649202, respond correctly to the effects of Cre recombinase.

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