Award details

Development and application of real time biosensors

Reference	BB/F004753/1
Principal Investigator / Supervisor	Professor Philip Poole
Co-Investigators / Co-Supervisors	Dr Victoria Bamford, Dr Alison East, Dr Ramakrishnan Karunakaran, Professor Helen Osborn, Professor Kimberly Watson
Institution	John Innes Centre
Department	Molecular Microbiology
Funding type	Research
Value (£)	971,949
Status	Completed
Type	Research Grant
Start date	01/06/2008
End date	31/05/2011
Duration	36 months

Abstract

Biosensors are powerful tools for detection and monitoring of compounds in the environment and in the chemical and pharmaceutical industries. They can be found in a number of forms including induction biosensors, electrical biosensors and FRET biosensors. These nanosensors operate at the atomic to microscopic/cellular level, providing the ability to detect the presence of compounds as well as in some cases rates of flux of compounds. Building on our isolation of a whole new suite of solute binding protein (SBP) induction biosensors, we now propose to develop a targeted group of these into FRET biosensors. FRET biosensors allow the real time monitoring of compounds in living cells and permit rates of flux to be measured. In particular we propose to express selected sugar, ion and pharmaceutically important SBPs and measure their ligand binding affinity. They will then be expressed as eCFP-SBP-eYFP fusion proteins for measurement and optimisation of their FRET signals. In addition 6-10 of the most novel of the SBPs will be crystallised to enable rational modification of their ligand specificity and FRET optimisation. In a radical new development we also propose to develop a targeted metagenomic library for the isolation of novel induction biosensors. This library will be composed of genomic DNA fragments from a selection of sequenced plant and environmental bacteria. We are confident such an approach can revolutionise the whole area of isolation of novel induction biosensors. In future work this technique could be applied to almost any compound of interest and provides extraordinary selective power.

Summary

Biosensors are devices of biological origin that can detect the presence of chemicals or other physical factors (e.g. light, temperature) in the environment. We are proposing to develop a whole new group of chemical biosensors, in particluar for sugars and simple ions like zinc, manganese and phosphate. These biosensors are examples of nanotechnology because they can sense chemicals at the atomic to cellular level. The particular biosensors we are proposing to develop are based on simple proteins called solute binding proteins. These proteins can be fused to two other proteins (CFP and YFP) to generate a fluorescent signal when they bind a specific chemical, which in this case will be a sugar or an ion. These nanosensors can be incorporated into living cells where they will report in real time on the concentration of the chemical they can sense. This is very important because it means changes in concentrations (flux) of chemicals can be measured. Many signalling mechanisms in biology depend on changes in the concentrations of metabolites so being able to sense these changes is crucial to many fundamental studies on biological action.

Committee	Closed Committee - Engineering & Biological Systems (EBS)
Research Topics	Microbiology, Plant Science, Structural Biology, Technology and Methods Development
Research Priority	X – Research Priority information not available
Research Initiative	Technology Development Initiative 2 (TDRI2) [2007]
Funding Scheme	X – not Funded via a specific Funding Scheme

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