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Title:
Synthetic Biology: design and construction of a steroid biosensor
Authors & affiliations:
Seppe Dierckx1, Wolfgang Eberle2, Jos Vanderleyden1
1
Centre of Microbial and Plant Genetics (CMPG), KULeuven, Belgium; 2Imec,
Belgium
Abstract: (Your abstract must use Normal style and must fit in this box. Your abstract should be no
longer than 300 words. The box will ‘expand’ over 2 pages as you add text/diagrams into it.)
The increased usage of oral contraceptives is becoming dangerous for our water safety.
In order to see how severe the danger is, an adequate sensor for steroids hormones is
neccesary since as of now, no in situ sensors are available. Therefore we had decided to
explore the possibilities to construct a steroid biosensor, based on the use of engineered
bacterial cells.
To develop the input module of the sensor we are making use of both the HpaA and the
RepA proteins, found in Escherichia coli and Comamonas testosteroni respectively.
HpaA detects and binds to phenylacetic acid, a plant hormone, while RepA binds to
human steroid sex hormones. Both act as transcription factors after binding their
respective ligands.
At first a proof-of-principle biosensor has been developed using the HpaA receptor.
When connected to a lacZ output module, HpaA has been found to elicit a linear doseresponsive result when induced by (hydroxy)phenylacetic acid, a prerequisite for an
adequate sensor.
The HpaA input module has also been connected to an electrical readout output module,
producing a compact, working biosensor. Data gathered by this construct will be used to
create a general in silico model of a hormone biosensor.
The human steroid biosensor makes use of the RepA receptor to detect testosterone and
various estrogens. Experiments have shown that when induced by these steroids RepA
produces a dose dependent response. Further constructions using RepA are yet to be
developed.
The aim of this project is to construct two working biosensors, one proof-of-principle
plant hormone biosensor and one human steroid hormone biosensor.
When combined with a suitable output module, for instance the electrical readout
module based on the production of pyocyanin, a compact in situ sensor can be
constructed . This could become a conventient tool to test water for their hormone
content.