Team:Michigan/Project

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Overall project

Our project has several different tracks that we will be working on in the wet-lab simultaneously. These tracks include quorum sensing, pili hyperexpression, and surface display. We made the decision to work in several tracks in order to maximize the efficiency of our lab teams. Our goal is to be able to combine all of these parts in the end and form an organism that will be able to effectively flocculate microalgae.

Quorum Sensing

Our cells will use quorum sensing to determine when the flocculation will start. The cells will produce an inducer either AHL or AI-2. You can find their lab notebook here.

Pili Hyperexpression

Type 1 pili (also known as fimbriae) are proteinaceous adhesins that are found on the surface of E. coli. One cell can contain up to 100 pili, which can form up to 2 um long. The pili help E. coli. form biofilms.

The pili are conrolled by the fim operon. This operon consists of several genes. The pili themselves are composed of several thousand subunits of FimA. The tip of each pili consists of the genes FimF, FimG, and FimH. FimH is an adhesin and is linked to FimA through FimF and FimG. Inside the cell, FimC carries proteins to the structural platform, FimD, which then assembles the pilus rod. This whole process is regulated by the genes FimB and FimE. These genes control an invertible DNA sequence, which, when in the "on" position, promote the production of pili.

By overproducing the pili, we hope to increase flocculation. By hyperexpressing FimH, we can increase the amount of pilus adhesin (Fig. 1), which will hopefully strengthen the E. coli binding to the algae. You can find their lab notebook here.

Fig. 1 C shows normal pili, B shows hyperexpressed pilus adhesin.

Virus Surface Display

Washboard