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ELECTROPHORESIS OF PROTEINS: DATA CAPTURE, ANALYSIS, AND CONSTRUCTION OF DATABANKS

Objective

IMPROVED AND MORE EFFICIENT TOOLS FOR THE SCREENING OF MICROBIAL STRAINS FOR BIOTECHNOLOGICAL PROCESSES, DETECTION OF CONTAMINANTS, IDENTIFICATION OF STRAINS, ESPECIALLY PATENTED BIOTECHNOLOGICAL AND PATHOGENIC STRAINS.

STUDY OF GENE EXPRESSION IN EUKARYOTE CELLS, UNDERSTANDING AND IMPROVED DIAGNOSIS OF DISEASE-RELATED PROTEIN ABNORMALITIES.
1-dimensional and 2-dimensional electrophoretic techniques provide powerful tools for the analysis of complex protein mixtures. Computer analysis of 1-dimensional and 2-dimensional protein patterns to establish databases of pathogenic bacteria, biotechnologically important bacteria and eucaryotic cells provides methods for screening and identifying bacterial strains, recognizing new groups, and characterizing human and animal disease related protein abnormalities. For wide use, standardization of techniques is of paramount importance.

A standard procedure has been developed for the production of 1-dimensional gels. A set of bacteria independently studied in 2 laboratories gave the same analytical result and the portability of the systems was demonstrated despite implementation on different computers using similar software. Databases were established for identification of bacterial species and protein types within species and standard 2-dimensional methodology was applied to human, murine, bacterial and streptomycete cells. A system using immobilized pH gradients for isoelectric focusing (IEF) was also developed providing greater reproducibility of 2-dimensional patterns. Computer systems of 2-dimensional gel analysis namely PDQUEST and BioImage were implemented and databases were established for the studied cell types. Standardized gels, analytical system and databases will allow major improvements in the study of developmental and disease processes in humans, and reliable, rapid identification of pathogenic and biotechnologically important bacteria.
USING A MASSCOMP MCS 5500 COMPUTER SYSTEM AND A PDQUEST SOFTWARE PACKAGE FOR THE ANALYSIS OF 2-D ELECTROPHORESIS DATA, PROTOTYPICAL DATA BANKS WILL BE CONSTRUCTED WITH LIMITED SETS OF BIOLOGICAL SAMPLES. THESE DATABANKS WILL HELP IN THE DESIGN OF IMPROVED SOFTWARE FOR USE WITH LARGE DATA SETS AND FOR THE DEVELOPMENT OF DEDICATED SYSTEMS WITH RAPID PROCESSORS.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

Institut Pasteur
Address
28 Rue Du Docteur Roux
75724 Paris
France

Participants (4)

Central Public Health Laboratory
United Kingdom
Address
61 Colindale Avenue
NW9 5HT London
JOYCE LOEBL LTD
United Kingdom
Address

Gateshead
RIJKSUNIVERSITEIT GENT
Belgium
Address

Gent
Royal Postgraduate Medical School
United Kingdom
Address
Du Cane Road
W12 0HS London