Difference between revisions of "Team:FSU/Results"

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Revision as of 22:02, 2 December 2018

Results

Although the Berkeley Test Cells showed an increase in fluorescence when exposed to sound. Further testing must be done to further characterize this initial response to sound. The Berkeley promoter BBa_K112402 was found to be a constitutive promoter. The same promoter, BBa_K112402, also shows a possible RFP upregulation when exposed to sound however further testing needs to be done to determine a more detailed relationship between amplitude, frequency and promoter upregulation. The FSU iGEM promoters: BamE, OsmC, and PspA also showed an increase in fluorescence after sound exposure.

In addition to characterizing the promoters proposed by UC Berkeley in 2008, we wanted to bring IGEM the BIOFAB collection of promoters. We picked three promoters, ApFAB90, ApFAB46, and ApFAB82. The activity of the promoters was characterized by inserting them into a PSCB1C3 plasmid which contained a red cassette that expresses RFP. We measured activity by using a ratio of Fluorescence to optical density and comparing initial and final values. We expected each promoter to behave with low, medium, and high activity respectively. The BIOFAB promoter behavior was not in the order that we were expecting. The apFAB46 showed the lowest activity but was expected to be the highest. ApFAB90 was found to have medium expression, however, was expected to be low activity. Finally, apFAB82 showed the highest fluorescence, however, was expected to be the medium expression. Further investigation is needed to determine these unexpected results.

Cells Mean RFU/OD700 Standard Deviation Mean RFU Standard Deviation Mean OD700 Standard Deviation
Negative Control 81 11.8 49 6.7 0.601 0.011
Positive Control 28642 2781.7 12028 854.2 0.425 0.074
BBa_K112400 12872 250.6 11993 425.0 0.932 0.026
BBa_K112401 2861 41.4 3082 164.7 1.077 0.057
BBa_K112402 24572 837.7 20455 507.1 0.833 0.014
BBa_K112405 304 11.3 246 12.6 0.807 0.013
BBa_K112407 148 0.5 131 2.5 0.882 0.016
PosmA 190 10.5 85 2.0 0.449 0.036
PpspA 828 46.2 371 9.7 0.449 0.030
PbamE 165 16.2 73 7.2 0.441 0.010
PzntA 168 26.1 73 5.6 0.439 0.036

Using a constitutive promoter as a positive control, we compared the flourescence/OD700 of the Berkeley promoters when allowed to grow in the incubator with “ambient sound”. We found that the expression of BBa_K112402 was comparable to he positive control. We believe this might be an indication that BBa_K112402 is a constitutive promoter. BBa_K112400 also had a high expression and we believe that this may indicate that it may also be a weaker constitutive promoter.



The BIOFAB promoters showed an unexpected level of activity. The promoter apFAB46 showed the lowest activity but was expected to be the highest. ApFAB90 was found to have medium expression, however, was expected to be low activity. Finally, apFAB82 showed the highest fluorescence, however, was expected to be the medium expression.



We found that two of the Berkeley promoters appeared to be constitutive promoters since they showed significant fluorescence after growing under ambient sound. The PSPA FSU system was likely slightly leaky since it shows a slight trace of fluorescence under ambient sound.



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Ambient_sound_results_graph.png

Ambient_sound_results.png

Ambient_sound_results2.png

Ambient_sound_results2_table.png

8kHz_80_dB_sound.png

8kHz_80_dB_data_table.png

8_kHz_100_dB_sound.png

8_kHz_100_dB_sound_data_table.png

2kHz_100_dB_sound.png

2_kHz_100_dB_data_table.png

2_kHz_100_dB_comparative_graph.png

2_kHz_100_dB_comparative_graph_data_table.png