Part:BBa_M36246:Experience

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Experience with BBa_M36246

We tested this part with the Gemini reporter. We found that the more cadmium we added, the less B-gal expression we had, and our GFP expression analysis gave us no conclusive results. We tested out construct along with a rhamnose induced Gemini strain, referenced as p82, and found that both produced the same result.

We ran an experiment of E.Coli strain 4-2 with varying concentrations of cadmium which showed that at concentrations of less than or equal 1mM, cell growth was not affected,. This taken into account, our results for p82 and our construct were quite interesting. Our ß-gal expression showed us a trend that we did not expect across both strains. Both p82 and our construct showed decreasing amounts of ß-gal proportional to increasing amounts of cadmium. Most interestingly, the 0M concentration has the most ß-gal expression of all our tests. Because our negative control was coming up with positive results, we have come to a few results. First, our construct is most likely not working. Although displaying the same trend, our construct has significantly less ß-gal expression that p82 alone. The graph shows the trend of increasing ß-gal expression as cadmium decreases across both bacteria strains. In order to compare them on the same scale, we divided the values by the 0M expression value. The trend was opposite of what we expected. We should have seen an increased amount of expression due to increased amounts of cadmium, not the opposite. Finally, we feel there is something wrong with the actuator we used. Our negative control was flawed, and thus we cannot gauge if our construct did anything in terms of ß-gal expression in the cells. When looking at GFP expression, our data was inconclusive, there was no trend in data across any of the strains.

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