Difference between revisions of "Part:BBa K2235003:Experience"
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At first, we have measured the cell viability of TOP10 in different concentrations of cumate over time. Our aim was to determine whether cumate is cytotoxic or not, before we measure the fluorescence. | At first, we have measured the cell viability of TOP10 in different concentrations of cumate over time. Our aim was to determine whether cumate is cytotoxic or not, before we measure the fluorescence. | ||
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The growth curve clearly demonstrates that cumate does not affect the cellular viability and proliferation of our strain. | The growth curve clearly demonstrates that cumate does not affect the cellular viability and proliferation of our strain. | ||
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Next, we employed the same concentrations and measured the fluorescence over time. We also used a negative control (transformed bacteria which is not expressing BFP). | Next, we employed the same concentrations and measured the fluorescence over time. We also used a negative control (transformed bacteria which is not expressing BFP). | ||
| − | [[Image:Expression of BFP in cumate. | + | [[Image:Expression of BFP in cumate.jpg|200px|thumb|left|]] |
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The fluorescence intensity is higher in all samples, even the untreated one compared to the negative control. Therefore, there is expression of BFP but not in a dose dependent manner. Normally, we would expect to see fluorescence only in the treated samples since the cumate binds to the repressor enabling the transcription of the downstream gene. However, we assume that the cymR is mutated at a specific site based on the sequencing results; thus, the expressed protein is unable to bind to the T5 operon in the first place and suppress the expression of BFP. | The fluorescence intensity is higher in all samples, even the untreated one compared to the negative control. Therefore, there is expression of BFP but not in a dose dependent manner. Normally, we would expect to see fluorescence only in the treated samples since the cumate binds to the repressor enabling the transcription of the downstream gene. However, we assume that the cymR is mutated at a specific site based on the sequencing results; thus, the expressed protein is unable to bind to the T5 operon in the first place and suppress the expression of BFP. | ||
Revision as of 13:53, 23 October 2017
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Applications of BBa_K2235003
At first, we have measured the cell viability of TOP10 in different concentrations of cumate over time. Our aim was to determine whether cumate is cytotoxic or not, before we measure the fluorescence.
The growth curve clearly demonstrates that cumate does not affect the cellular viability and proliferation of our strain.
Next, we employed the same concentrations and measured the fluorescence over time. We also used a negative control (transformed bacteria which is not expressing BFP).
The fluorescence intensity is higher in all samples, even the untreated one compared to the negative control. Therefore, there is expression of BFP but not in a dose dependent manner. Normally, we would expect to see fluorescence only in the treated samples since the cumate binds to the repressor enabling the transcription of the downstream gene. However, we assume that the cymR is mutated at a specific site based on the sequencing results; thus, the expressed protein is unable to bind to the T5 operon in the first place and suppress the expression of BFP.
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