Award details

Improving archaeal DNA polymerases for molecular biology applications

Reference	E19804
Principal Investigator / Supervisor	Professor Bernard Connolly
Co-Investigators / Co-Supervisors
Institution	Newcastle University
Department	Inst for Cell and Molecular Biosciences
Funding type	Research
Value (£)	222,497
Status	Completed
Type	Research Grant
Start date	01/03/2004
End date	28/02/2007
Duration	36 months

Abstract

Archaeal DNA polymerases are unable to replicate template strand uracil and incorporating this base into DNA (e.g. as a dUTP contaminant of the dNTP mixture; by deamination of either dCTP to dUTP or G:C base-pairs to G:U mispairs), during PCR, results in complete inhibition. These enzymes, therefore, often show inefficient PCR performance and cannot be used in contamination-free applications where dUTP replaces TTP. Archaeal polymerases contain a uracil binding pocket, responsible for uracil sensitivity, which will be inactivated by mutagenesis. Random mutagenesis will be used to improve processivity and confer reverse transcriptase activity. These altered polymerases should show superior performance in a number of PCR applications. The mutations will be fully characterised to elucidate mechanistic features and bench-marked in a number of real PCR applications.

Summary

unavailable

Committee	Closed Committee - Engineering & Biological Systems (EBS)
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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