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Molecular analysis of the enzymic properties of human poly (ADP-ribose) polymerase using random and site-directed mutagenesis

ReferenceC05217
Principal Investigator / Supervisor Dr Minnie O'Farrell
Co-Investigators /
Co-Supervisors
Institution University of Essex
DepartmentBiological Sciences
Funding typeResearch
Value (£) 155,697
StatusCompleted
TypeResearch Grant
Start date 01/04/1996
End date 01/04/1999
Duration36 months

Abstract

This study uses mutagenesis of the cDNA to make a detailed molecular analysis of the structure and enzymology of the nuclear enzyme, poly (ADP-ribose) polymerase. The catalytic domain of the enzyme will be subject to random mutagenesis using error-prone PCR and loss of function and gain of function mutants will be isolated. Mutants already isolated following chemical mutagenesis will be further explored using site-directed mutagenesis. All the mutants isolated will be sequenced and characterised by estimating Km, Vmax and the kinetics and structure of polymer product formed. Selected mutants will be taken forward for more detailed analysis in an in vitro assay to explore DNA binding, protein-protein interactions and inter- and intramolecular modifications by the enzyme.

Summary

unavailable
Committee Closed Committee - Biochemistry & Cell Biology (BCB)
Research TopicsX – not assigned to a current Research Topic
Research PriorityX – Research Priority information not available
Research Initiative X - not in an Initiative
Funding SchemeX – not Funded via a specific Funding Scheme
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