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Directed evolution of cystatins for enhanced resistance to nematodes: off-rate versus specificity spectrum
Reference
G15882
Principal Investigator / Supervisor
Professor Michael McPherson
Co-Investigators /
Co-Supervisors
Institution
University of Leeds
Department
Plant Biochemistry & Biotechnology
Funding type
Research
Value (£)
201,772
Status
Completed
Type
Research Grant
Start date
01/04/2002
End date
31/03/2005
Duration
36 months
Abstract
Cystatins are small protein inhibitors of cysteine proteases that have proven useful as a basis for transgenic resistance against nematodes that cause crop damage. Achieving economically viable levels of resistance requires further improvement of the system. One approach is to select plant-based cystatins with decreased off-rates. We have used DNA shuffling of several plant cystatin genes to build a phage display library of cystatins. This project will screen the library against various cysteine proteases to select improved variants. Surface Plasmon Resonance will be used to characterise the relative binding properties of the cystatins to a range of target proteases, including expressed plant nematode proteases. The relationship between decreased off-rate and retention of broad- spectrum protease specificity will be examined and structural studies initiated.
Summary
unavailable
Committee
Closed Committee - Genes & Developmental Biology (GDB)
Research Topics
X – not assigned to a current Research Topic
Research Priority
X – Research Priority information not available
Research Initiative
X - not in an Initiative
Funding Scheme
X – not Funded via a specific Funding Scheme
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