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EUROPEAN RESOURCE CENTRES FOR PLASMID BEARING BACTERIAL STRAINS

Objective

THE ACADEMIC AND INDUSTRIAL SCIENTIFIC COMMUNITY WILL BE PROVIDED WITH A STOCK OF MEDICALLY IMPORTANT PLASMID BEARING STRAINS FOR RESEARCH AND PRODUCT DEVELOPMENT.

THIS SERVICE WILL BE MADE WIDELY KNOWN, THROUGH THE CATALOGUE OF THE COLLECTION, PUBLIC HEALTH LABORATORY SERVICE PUBLICATIONS OFFICER, EUROPEAN CULTURE COLLECTION CURATORS ORGANIZATION (ECCCO), WORLD FEDERATION FOR CULTURE COLLECTIONS (WFCC), AND OTHER AVENUES.
A collection of plasmid bearing bacteria has been established, as freeze dried ampoule stocks, which is quality controlled and catalogued (computer database). This specialised resource is unique and the clinical and scientific community has already made extensive use of the collection. Molecular biology laboratory techniques have been used for quality control.

European culture collections have lacked centres with the specialized skills necessary to handle plasmid bearing strains, and thus there were no provisions for their collection, preservation and distribution. The aim was to establish centres with quality control procedures suitable for collections of plasmid bearing strains of biotechnological and medical importance, including patent strains.

Methods and quality control procedures were developed which allow the freeze drying of plasmid bearing strains without loss or alteration of their characteristics. The means of quality control established include: before and after freeze drying checks on stability of host cells, plasmid presence and absence, molecular weight of plasmids, confirmation of phenotypic markers and, where appropriate, restriction digests of plasmid deoxyribonucleic acid (DNA).
Seed stocks for rebatching are kept in liquid nitrogen, and plasmid bearing strains are distributed in freeze dried ampoules.
The databases established at each collection are an information resource and greatly facilitate the production of catalogues and searches.
TO COLLECT, PRESERVE, QUALITY-CONTROL, AND MAKE AVAILABLE TO THE SCIENTIFIC COMMUNITY (ACADEMIC AND INDUSTRIAL) MEDICALLY IMPORTANT PLASMID BEARING BACTERIA STRAINS.

TO LAY DOWN STOCKS OF COLLECTED STRAINS, AS AGAR SLOPES, AS FREEZE-DRIED AMPOULES, AND IN LIQUID NITROGEN; TO COMPARE THESE DIFFERENT METHODS OF PRESERVATION TO DETERMINE THE OPTIMUM METHOD.

TO CATALOGUE COLLECTED STRAINS AND ENTER DATA INTO A MICROCOMPUTER. TO SET UP ROUTINE QUALITY CONTROL PROCEDURES FOR PRESERVED PLASMIDS AND THEIR PROPERTIES : BY ELECTROPHORESIS/CENTRIFUGATION DETERMINE PRESENCE/ABSENCE OF PLASMIDS BEFORE/AFTER PRESERVATION; MOLECULAR WEIGHTS; RESTRICTION ENDONUCLEASE PATTERNS, COPY NUMBERS. TO QUALITY CONTROL THE GENETIC EXPRESSION OF PLASMID CODED PROPERTIES (E.G. ANTIBIOTIC RESISTANCES, TOXIN PRODUCTION, ETC) AND SUCH PROPERTIES AS REPLICATION, TRANSMISSIBILITY.

TO INVESTIGATE THE FEASIBILITY OF PRESERVING PLASMIDS ALONE, I.E. AS DNA.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

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

Participants (1)

Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH
Germany
Address

37001 Göttingen