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Results
E. coli Environment Alters Tumbling:
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Salt Experiment
CONTROL
0.0 M NaCl
| RPS (rotations/sec) | 0.21 | 0.17 | 0.48 | 0.71 | 0.45 |
| Frequency (Hz) | 132 | 107 | 302 | 446 | 283 |
0.5 M NaCl
| RPS (rotations/sec) | 0.80 | 0.83 | 0.72 | 0.71 | 0.71 |
| Frequency (Hz) | 503 | 522 | 452 | 446 | 446 |
0.9 M NaCl
| RPS (rotations/sec) | 0.21 | 0.55 | 0.71 | 0.40 | 0.54 |
| Frequency (Hz) | 132 | 346 | 446 | 251 | 339 |
CheY
0.0 M NaCl
| RPS (rotations/sec) | 0.57 | 0.60 | 0.68 | 0.78 | 0.74 |
| Frequency (Hz) | 358 | 377 | 427 | 490 | 465 |
0.5 M NaCl
| RPS (rotations/sec) | 0.32 | 0.40 | 0.43 | 0.25 | 0.35 |
| Frequency (Hz) | 201 | 251 | 270 | 157 | 220 |
0.9 M NaCl
| RPS (rotations/sec) | 0.31 | 0.32 | 0.21 | 0.25 | 0.28 |
| Frequency (Hz) | 195 | 201 | 132 | 157 | 176 |
CheZ
0.9 M NaCl
| RPS (rotations/sec) | 0.63 | 0.50 | 0.25 | 0.63 | 0.31 |
| Frequency (Hz) | 396 | 314 | 157 | 395 | 195 |
CheY vs. CONTROL
| Salt Concentration | 0.0M NaCl | 0.5M NaCl | 0.9M Nacl | |||
| Control | CheY | Control | CheY | Control | CheY | |
| Avgerage frequency (Hz) |
254 |
424 |
474 |
220 |
303 |
172 |
| Standard Deviation |
138 |
56 |
36 |
44 |
118 |
28 |
| Two-tailed t-test (a=0.05) |
0.0127 |
0.0003 |
0.0978 |
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CheY vs. CheZ vs. CONTROL
| Control | cheY | cheZ | |
| Avgerage frequency (Hz) | 303 | 172 | 292 |
| Standard Deviation | 28 | 118 | 115 |
| Two-tailed t-test (a=0.05) | 0.0978 | 0.9136 |
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Inducible Tumbling System in cheY and cheZ
In order to test our bacterial communication system, we transformed cheY and cheZ parts into E. coli B strain HT115. Our BioBricks included an inducible T7 promoter that is activated by IPTG. As shown in the figure, 100% of successfully transformed bacteria with the T7 RBS cheY construct showed tumbling, demonstrating the functionality of the T7 RBS cheY part.
Reconstitution of Flagella
Our team created the following BioBricks that control the assembly and operon regulation of E. coli flagella: BBa_K842000 flhB, BBa_K842001 flhD, BBa_K842002 flgJ, BBa_842003 fliA, BBa_K842005 fliH, BBa_K842011 motA, BBa_K842012 motB. These parts allow for the reconstitution of a flagella mechanism in E. coli B strains where the flagella mechanisms are lacking.
Future Directions
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pH Experiment
CONTROL
pH 5
| RPS (rotations/sec) | 0.25 | 0.33 | 0.36 | 0.20 | 0.42 |
| Frequency (Hz) | 157 | 207 | 226 | 126 | 264 |
pH 6
| RPS (rotations/sec) | 0.43 | 0.36 | 0.46 | 0.40 | 0.25 |
| Frequency (Hz) | 270 | 226 | 289 | 251 | 157 |
pH 7
| RPS (rotations/sec) | 0.21 | 0.17 | 0.48 | 0.71 | 0.45 |
| Frequency (Hz) | 132 | 107 | 302 | 446 | 283 |
pH 8
| RPS (rotations/sec) | 0.25 | 0.50 | 0.25 | 0.35 | 0.33 |
| Frequency (Hz) | 157 | 314 | 157 | 220 | 207 |
pH 9
| RPS (rotations/sec) | 0.26 | 0.25 | 0.33 | 0.33 | 0.32 |
| Frequency (Hz) | 163 | 157 | 207 | 207 | 201 |
| pH | 5 | 6 | 7 | 8 | 9 |
| Average frequency (Hz) | 196 | 238 | 254 | 211 | 187 |
| Standard Deviation | 55 | 51 | 138 | 64 | 24 |
