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Contenu archivé le 2024-04-15

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

Objectif

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.
A standard procedure for the production of improved 1-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoretic (SDS-PAGE) protein profiles of pathogenic bacteria has been developed. Software for the analysis and comparison of such profiles for identification and characterisation purposes have been implemented. Full system portability between laboratories has been demonstrated. Work has been further extended to 2-dimensional separation of proteins.

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.
SOFTWARE AND DATABANK FOR 1-D PATTERNS OF PATHOGENIC BACTERIA HELD IN THE NCTC AS A RAPID TECHNIQUE FOR IDENTIFICATION AND TYPING AND TO SELECT STRAINS FOR 2-D PAGE. COLLABORATION WITH THE UNIVERSITY OF GHENT IN STANDARDIZING GEL RUNNING PROCEDURES; ANALYSIS OF 2-D GELS IN COLLABORATION WHITH THE ROYAL POSTGRADUATE MEDICAL SCHOOL, LONDON. FOR 2-D WORK, COMPARISON OF MEMBRANE, RIBOSOMAL AND WHOLE-CELL PROTEINS WILL BE USED TO STUDY MICROBIAL RELATIONSHIPS. NCTC DATA WILL BE INCLUDED IN THE PROPERTY DATABANK OF THE UK DEPT. OF TRADE AND INDUSTRY (MICIS) AND OTHER APPROPRIATE EUROPEAN DATABANKS (EG MINE).

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Central Public Health Laboratory
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Adresse
61 Colindale Avenue
NW9 5HT London
Royaume-Uni

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