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

Información del proyecto

Identificador del acuerdo de subvención: BAP*0138

Proyecto cerrado

Fecha de inicio 1 Julio 1986

Fecha de finalización 30 Abril 1991

Financiado con arreglo a

• Multiannual research action programme (EEC) in the field of biotechnology (BAP), 1985-1989

Coste total

Sin datos

Aportación de la UE

Sin datos

Coordinado por

RIJKSUNIVERSITEIT GENT
Belgium

Objetivo

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.
The technique for one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of proteins was standardised to enhance reproducibility, necessary for computer comparison, database construction and portability. The process was completely tested and the reproducibility obtained was sufficient to allow the possibility of large and long term database construction.

A Siemens 7570-C implemented FORTRAN 77 and UCSD-like PASCAL program has been developed for the storage, retrieval and modification of one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) protein profiles. The database program was extended with procedures for both fast identification and numerical comparison of a limited set of 1-dimensional SDS-PAGE protein profiles (up to 800).

A microcomputer implemented PASCAL program was developed for the automated alignment of 1-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) protein profiles. The electrophoretic pattern of bacterial strain Psychrobacter immobilis LMG 1125 is used as a universal reference profile.

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.
DEVELOPMENT OF SOFTWARE AND DATABANK FOR 1-D PATTERNS OF BIOTECHNOLOGICALLY IMPORTANT BACTERIA, USING STRAINS HELD IN THE LABORATORY OF MICROBIOLOGY, GENT CULTURE COLLECTION (LMG), ON WHICH EXTENSIVE GENOMIC AND BIOTECHNOLOGICAL DATA EXISTS. THERE WILL BE CLOSE COLLABORATION WITH THE NATIONAL COLLECTION OF TYPE CULTURES (NCTC).COLINDALE, UK ON PREPARATION AND RUNNING OF GELS, EXCHANGE OF SOFTWARE FOR HARDWARE COMMON TO BOTH (LKB DENSITOMETERS/APPLE MICROCOMPUTERS), EXCHANGE OF STRAINS, EXTRACTS AND GELS. 2-D PAGE ON SELECTED STRAINS WILL BE ANALYSED IN COLLABORATION WITH THE ROYAL POSTGRADUATE SCHOOL, LONDON AND INSTITUTE PASTEUR, PARIS.

Ámbito científico (EuroSciVoc)

• ciencias naturales › informática y ciencias de la información › base de datos
• ciencias naturales › ciencias biológicas › microbiología › bacteriología
• ciencias naturales › ciencias químicas › química inorgánica › metales alcalinos
• ciencias naturales › ciencias biológicas › bioquímica › biomoléculas › proteínas
• ciencias naturales › ciencias químicas › electroquímica › electroforesis

Programa(s)

• FP1-BAP - Multiannual research action programme (EEC) in the field of biotechnology (BAP), 1985-1989

Tema(s)

Datos no disponibles

Convocatoria de propuestas

Datos no disponibles

Régimen de financiación

CSC - Cost-sharing contracts

Coordinador

RIJKSUNIVERSITEIT GENT

• Aportación de la UE: Sin datos
• Dirección: LAB.VOOR MICROBIOLOGIE LEDEGANCKSTRAAT 35 ; 9000 GENT ; Bélgica
• Coste total: Sin datos

Participantes (5)

AND WITH THE COLLABORATION OF:

• Aportación de la UE: Sin datos
• Coste total: Sin datos

Central Public Health Laboratory

Reino Unido

• Aportación de la UE: Sin datos
• Dirección: 61 Colindale Avenue ; NW9 5HT London
• Coste total: Sin datos

INSTITUT PASTEUR

Francia

• Aportación de la UE: Sin datos
• Dirección: Rue du Docteur Roux 25 ; 75724 PARIS
• Coste total: Sin datos

Joyce Loebl Ltd

Reino Unido

• Aportación de la UE: Sin datos
• Dirección: Dukesway Team Valley Trading Estate ; NE11 0PZ Gateshead
• Coste total: Sin datos

Royal Postgraduate Medical School

Reino Unido

• Aportación de la UE: Sin datos
• Dirección: Du Cane Road ; W12 0HS London
• Coste total: Sin datos