Team:Pittsburgh/Proof Of Concept

Bluetooth Bacteria

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Proof of Concept

 

Temperature Sensor

DnaK, when incubated with shaking, produces significant mWasabi expression. This simulates theconditions generated by the heating of MNPs as it would be localized around each cell, instead of through the container and media. DnaK produced the highest level of fluorescence and, unlike TlpA and TcI, consistently produced mWasabi expression throughout the cell culture. The shaking of the culture simulates thermal induction via MNPs which produced a detectable difference in mWasabi output.

Figure 1. DnaK increased mWasabi expression at 45C with shaking

Figure 2. High percentage of cells in culture fluorescing due to mWasabi

Magnetic Nanoparticle Attachment

Furthermore, we had successful attachment of the particles to our cells. The OD reading decreases after AF-MNPs were added to bacteria suspended in PBS and the nanoparticle-bacteria conjugate was removed with a magnet. If the mass removed by the magnet only contained the AF-MNPs, the OD reading should stay around the same. If the mass removed by the magnetic contains nanoparticle-bacteria conjugate. Since the OD decreased, it suggests that nanoparticle-bacteria conjugates were being formed. The formation of nanoparticle-bacteria conjugates implies that the AF-MNPs were localized around the bacteria.

Figure 3. The nanoparticle-bacteria conjugate is attracted to the magnet

Table 1. OD change before and after the addition of MNPs

	Initial OD600 (no MNP added)	Final OD600 (MNP added and removed)
DH5alpha	Bba_J36848	LB + no antibiotic
Sample 1	7.006	5.874
Sample 2	1.886	0.561
Sample 3	0.411	0.608
Sample 4 (no cells, control)	0.202 (blank)	0.209 (blank)

Device

We have shown that our temperature sensors do allow gene expression at elevated temperatures and that our MNPs localize around the bacteria. The last step to our project is to build the AMF-producing device and demonstrate that gene expression can be induced by AMF. Although we were not able to build the device due to limited resources, we developed a detailed list of the materials that we would need as well as the circuit that we need to build with these parts based on our discussion with experts.

Figure 4. Circuit for the AMF-producing device