Difference between revisions of "Part:BBa K2235003:Experience"
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===Applications of BBa_K2235003===
 
===Applications of BBa_K2235003===
  
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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.
 
  
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[[Image:OD600 cumate.png|200px|thumb|left|]]
 
  
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[[Image:OD600 cumate.png|600px|thumb|left|Figure 1]]
  
  
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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.
  
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The growth curve clearly demonstrates that cumate does not affect the cellular viability and proliferation of our strain.
  
  
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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).
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[[Image:Expression of BFP in cumate.jpg|600px|thumb|left|Figure 2]]
  
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[[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.
 
  
  
  
 +
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 compared to the negative control. Therefore, the expression of BFP is not in a dose dependent manner since fluorescence can be detected even in the untreated sample. 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 suggest 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.
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 15:36, 25 October 2017


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Please enter how you used this part and how it worked out.

Applications of BBa_K2235003

Figure 1


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.







Figure 2




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 compared to the negative control. Therefore, the expression of BFP is not in a dose dependent manner since fluorescence can be detected even in the untreated sample. 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 suggest 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.

User Reviews

UNIQ530642136c9095b0-partinfo-00000000-QINU UNIQ530642136c9095b0-partinfo-00000001-QINU