Part:BBa_K1140006:Experience
Applications of BBa_K1140006
The team observed that this part works correctly in E. coli K12.
Surprisingly, we obtained different behaviors in clones transformed with the same DNA (figure 3). All measurements were performed at least in triplicate, the aritmethic mean is shown. For each measure in a given temperature, the system was left until a point in which we were sure the O.D of the cell culture and the production of the protein were in equilibrium, steady, and uniform, before the cells population started to decrease (which we found was 17h). We took as a standard for the RFUs the amount of fluorescence emitted by an E. coli K12 culture transformed with a constitutively expressed part BBa_E1010 (the amount of fluorescence emitted by our culture was calculated by dividing the fluorescence of the sample by the fluorescence of the standard). Figure 2 shows the behavior of our best clone, dubbed M1. M12 clone, showing a weird behavior, is to be sequenced to verify if this outcome is due to mutation or intrinsic cellular noise.
Mathematically, we found that a simple gaussian function fits our data well, and it provides us a way to quantify the strength (amplitude), optimal value (horizontal shift), and definition or clearness (width) of our RNAT activity (figure 4). We believe positive slope is due to RNAT melting, while negative slope is due to increase in the overall protein degradation rate due to higher temperatures. This function also allows for comparisons between different RNAT, as well as being potentially predictive for non verified temperatures.
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