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Proteomic analysis of oil-synthesising subdomains of the endoplasmic reticulum

Reference	JE412658
Principal Investigator / Supervisor	Professor Johnathan Napier
Co-Investigators / Co-Supervisors	Professor Michael Beale, Professor Peter Shewry
Institution	Rothamsted Research
Department	Plant Pathology and Microbiology
Funding type	Research
Value (£)	110,948
Status	Completed
Type	Research Grant
Start date	01/06/2000
End date	01/12/2000
Duration	6 months

Abstract

The synthesis of storage triacylglycerols and their assembly into oil bodies occurs in distinct sub-domains of the endoplasmic reticulum, catalysed by multi-enzyme complexes. Despite considerable effort many of the known enzymes involved in these biologically and economically important processes have not been purified or characterised, neither has it proved possible to identify corresponding cDNA or genes. We aim to use a different approach, combining the transformation of Arabidopsis and Brassica napus with genes for fluorescent (GFP)-tagged desaturase and oleosin, known components of the 'metabolome' involved in TAG synthesis. The use of spectrally distinct fluorescent tags will be used to prepare ER-sub-domain fractions which will be then subject to proteomic analysis of component proteins. This will allow the identification of corresponding EST or genomic sequences from the Arabidopsis genome programme. A MALDI-TOF mass spectrometer is required to carry out this work while the experimental approach will be applied in the future to the isolation and proteomic analysis of other sub-cellular fractions of interest.

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	Joint Equipment Initiative 1999 (JE4) [1999]
Funding Scheme	X – not Funded via a specific Funding Scheme

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