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GENETICS AND MOLECULAR BIOLOGY OF STREPTOMYCES : MAPPING, PLASMIDS INSTABILITY AND MLS-RESISTANCE.

Objective

BENEFITS STEMMING FROM THE COMPARATIVE STUDIES OF SEVERAL MACROLIDE PRODUCING STRAINS WILL BE:

THE ELUCIDATION OF THE MECHANISMS LEADING TO AMPLIFICATION OF DIFFERENT CLASSES OF AMPLIFIABLE UNITS.
THE USE OF THESE UNITS TO CONSTRUCT DIFFERENT INTEGRATIVE VECTORS WHICH WILL UNDERGO AMPLIFICATION WITHIN DIFFERENT MACROLIDE PRODUCERS.
THE FUTURE USE OF THESE VECTORS TO AMPLIFY GENES CODING FOR ANTIBIOTIC PRODUCTION, ANTIBIOTIC MODIFICATION, EXTRACELLAR ENZYMES, ENZYME INHIBITORS AND OTHER INTERESTING PRODUCTS.
THE CONSTRUCTION OF STRAINS STABLY PRODUCING THEIR SECONDARY METABOLITES.

THE OBJECTIVE OF THIS STUDY IS TO ELUCIDATE THE MOLECULAR EVENTS LEADING TO THE HIGH DEGREE OF INSTABILITY AND TO CHARACTERIZE UNSTABLE GENES SUCH AS THOSE CODING FOR RESISTANCE AGAINST MACROLIDES, OR FOR THE PRODUCTION OF MACROLIDES.

MACROLIDES RESISTANCE: IT WILL BE TESTED WHETHER THE INDUCIBLE RESISTANCE AGAINST DIFFERENT MACROLIDES IS DUE TO METYLATIONOF THE 23S RNA AND (OR) ANOTHER MECHANISMS. THE DIFFERENT CLONED MACROLIDE RESISTANCE GENES OF S.RETICULI WILL BE COMPARED TO THOSE OF S.AMBOFACIENS.

GENETIC INSTABILITY: IT WILL BE STUDIED THE MECHANISMS OF GENETIC INSTABILITY, BY MEANS OF PLASMIDS ISOLATED FROM STREPTOMYCES STRAINS PRODUCING MACROLIDES, AND ALSO FROM PLASMID-FREE STRAINS, IN ORDER TO STUDY THE CHROMOSOMAL DNA ALTERATIONS OCCURED BY DELETING AND/OR AMPLIFICATIONS.

INSERTION ELEMENTS: THE STRAINS THAT LOSE REVERSIBLY CERTAIN TRAITS WILL BE TESTED FOR THE PRESENCE OF INSERTION SEQUENCES WITHIN THESE TRAITS.

REPLICATION ORIGINS: WILL BE MAPPED, BECAUSE IT IS PARTICULAR INTEREST TO LOCATE THE REPLICATION ORIGINS WITH RESPECT TO THE REFERENCES WHICH UNDERGO AMPLIFICATION.

AMPLIFIABLE INTEGRATIVE VECTORS: AMPLIFIABLE UNITS OR PARTS OF THESE WILL BE CLONED INTO VECTORS NOT REPLICATING IN STREPTOMYCES, BUT CARRYING AN ANTIBIOTIC RESISTANCE GENE WHICH WILL BE EXPRESSED.

Coordinator

LUDWIG-MAXIMILIANS UNIVERSITY OF MUNICH
Address
Maria-ward-strasse 1A
80638 Muenchen
Germany

Participants (1)

Université de Paris XI (Université Paris-Sud)
France
Address

Guerineau