Team:Pittsburgh/Engineering

Bluetooth Bacteria

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Engineering Success

 

 

In the short lab time made available to the team, a design cycle attempted to carry out was based on an imagined concept of a system that would induce gene expression in E. coli wirelessly. After a considerable amount of baseline information gathered from literature, and virtual conceptual development, the team was able to morph this idea into a full design: alternating magnetic fields, produced from a device, would be utilized to trigger a localized heating response from magnetic nanoparticles bound to the surface of E. coli. Within E. coli, the localized heating would activate temperature-sensitive dimers, initiating gene expression, made visible by fluorescent protein mWasabi.

Progress and results were recorded in small successes for the team, such as the successful transformation of temperature-sensitive plasmids pTlpA-Wasabi, pTlpA-Wasabi-NF, pDnaK-Wasabi, pTcI-Wasabi, pRpoH-Wasabi, pLaqI-Wasabi, pClp-Wasabi, PHtpG-Wasabi, and pGrpE-Wasabi, from the Mikhail Shapiro Lab, into BL21 and DH5alpha competent cells. These transformed plasmids became crucial for the team’s ability to conduct thorough heat assay experimentation. From the evidence presented on the Results page, observed fluorescence recorded through the conducted heat assays were induced from localized heating provided by controlled temperature environments, suggesting that heating theoretically produced from magnetic nanoparticles should be sufficient for the initiation of gene expression.

In addition to these findings, the team also managed to conjugate amine-functionalized magnetic nanoparticles (AF-MNPs) to the surface of the bacteria, suggested from experimentation found on the Results page. We observed that the OD reading of bacteria suspended in PBS decreased after the AF-MNPs were added and the nanoparticle-bacteria conjugate was removed; this suggested that the AF-MNPs were localized around the bacteria surface, which made the bacteria respond the magnet. If the AF-MNPs did not attach to the bacteria surface through electrostatic interaction, the OD reading should remain the same because the mass removed by the magnet would not contain bacteria.

Furthermore, we implemented plasmids with constitutive RFP expression to test the relative mWasabi expression levels. For these constructs, each cell will both constitutively express RFP and express mWasabi in response to thermal induction. This allows us to measure levels of fluorescence using a plate reader, microscopy, flow cytometry. Experiments were performed in triplicate to verify validity. This can be found on the Results Page.