Award details

Developing stable transfection of Eimeria tenella

Reference	BBS/E/I/00001142
Principal Investigator / Supervisor	Professor Fiona Tomley
Co-Investigators / Co-Supervisors
Institution	The Pirbright Institute
Department	The Pirbright Institute Department
Funding type	Research
Value (£)	639,808
Status	Completed
Type	Institute Project
Start date	01/04/2003
End date	30/06/2009
Duration	75 months

Abstract

The development of stable transfection for a small number of protozoan pathogens is contributing greatly to understanding gene function in these organisms. Transient transfection of E. tenella sporozoites was established in our lab some years ago, but stable integration and expression of transgenes remains elusive. There are a number of reasons for this including low efficiency of transfection, the need to propagate transfected parasites in vivo by introduction of labile, electroporated, sporozoites into the avian host, a lack of well-defined and characterised promoter sequences, limited knowledge of the frequency with which parasites undergo non-homologous and homologous recombination and a lack of assays for detecting transgene integration and expression. The objectives of this project are i) Improve the efficiency of electroporation of plasmids into sporozoites (ii) Establish reliable infection of chickens with electroporated sporozoites (iii) Evaluate a range of Eimeria promoters for their abilities to drive transgene expression (iv) Express and evaluate additional selectable markers in vivo (v) Develop PCR-based assays for detection of integrated transgenes within parasites (vi) Design plasmid vectors suitable for targeted integration of transgenes via homologous recombination and (vi) Generate and propagate genetically altered parasites through the entire endogenous life-cycle.

Summary

unavailable

Committee	Closed Committee - Animal Sciences (AS)
Research Topics	Animal Health, Microbiology, Technology and Methods Development
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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