Team:Berkeley

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In iGEM, designing a bacterial biosensor is a well trodden path. From heavy metal detectors to rotting meat sensors, a variety of substrates have been detected in the past, prompting a colorimetric response via the synthesis of a pigment or fluorescent protein. However, many of these sensors depend on the transcription of a colorimetric reporter after a signal is detected. This means that upon signal reception, the bacterial cell must begin synthesis of proteins and enzymes responsible for the visible response - a process that can take upwards of 12 hours to produce a color change distinguishable by the naked eye. We think its time to revolutionize the traditional iGEM biosensor and make it considerably faster.



Cue the Indigo Biosensor. When the cellular system we are designing receives a signal, it will activate an enzyme involved in the indigo biosynthesis pathway responsible for quickly generating the ubiquitous indigo dye. This activated enzyme, a beta-glucosidase, will act on a glycosylated indigo derivative called indican that we are engineering to be constitutively produced by our cell. Indican is commonly produced in plant cells and is both soluble and colorless in the cytosolic environment. Upon activation of our beta-glucosidase, stores of colorless indican built up as our cell cultures proliferate will be efficiently and rapidly converted to blue indigo dye. Thus, our reporter system bypasses the time consuming process of transcribing and translating a reporter enzyme after a signal is detected and instead exploits a single enzyme's activity in going from no color, to a concentrated blue pigment.

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