Team:LambertGA
Characterization of Nonlysosomal Proteolysis
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Tuning Biosensors in a Switch-like Fashion
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Tuning Biosensors in a Switch-like Fashion
Tuning Biosensors in a Switch-like Fashion
Three million children die every year as a result of micronutrient deficiencies. The World Health Organization estimates that over 2 billion people worldwide are at risk and the Center for Disease Control estimates the costs of testing to be in the millions of dollars. Simple to use, inexpensive biosensors are an area of great interest in synthetic biology. Quite simply they could save millions of lives by providing inexpensive options to address micronutrient deficiencies while saving millions of dollars. The focus of this year’s Lambert’s iGEM team was to research the issues within the area of biosensors and engineer cost effective solutions for both scientists engineering the biosensors and the end users.
Biosensors often rely on the response of selection promoters to turn on or off protein expression. The concentration of proteins in a cell is determined by both the amount synthesized and the amount degraded. Thus, protein degradation is a crucial aspect of maintaining intramolecular equilibrium. A class of ATPases known as AAA+ Proteins involves a well-known proteolysis mechanism known as ClpXP in which ClpX unfolds and translocates a tagged protein into a sequestered proteolytic compartment in ClpP.
We engineered an inducible construct in which ClpXP will degrade a chromoprotein upon induction. Our system will first be tested in well characterized using Green Fluorescent Protein to establish baseline data. Chromo-protein expression data will be gathered using a device that can quantify the color of the light reflected by the chromoprotein before and after induction. This will ultimately allow us to measure the relative strength of degradation and further characterize a well-known proteolysis mechanism. Our characterization of ClpXP will serve as a precursor for controlled protein delivery in a switch for biosensors.
