Objectif DEVELOPMENT OF THE POSSIBILITY TO CLONE FOREIGN GENES INTO TWO OF THE MOST IMPORTANT INDUSTRIAL FUNGI : ASPERGILLUS ORYZAE USED FOR PRODUCTION OF CITRIC ACID AND PENICILLIUM CHRYSOGENUM USED FOR PRODUCTION OF PENICILLIN. IN PARTICULAR, THE FEASIBILITY OF USING THE NITRATE CONTROL REGION FOR INDUCTION OF THE EXPRESSION OF THE INSERTED INTERLEUKOCIDIN GENE WOULD BE OF PARAMOUNT COMMERCIAL INTEREST. A study was made fundamental and applied geneticaspects of filamentous fungi involving the development of genetic transformation and expression systems which are of central importance for the study and exploitation of filamentous fungi. Studies focussed on 2 selection systems which have a concomitant phenotype (ie, resistance to an antibiotic agent resulting in a specific growth defect). Certain acceptable fungi were programmed to synthesize and secrete heterologous proteins of commercial value in copious amounts.Transformation systems were successfully generated for a variety of industrial fungi based on the nitrate reductase system (niaD). These include Aspergillus oryzae, A niger, Penicillium chrysogenum, Gibberella fujikuroi, Cephalosporium acremonium. Transformation based on the uracil system (pyrG) was also developed for A oryzae and A niger. Such results indicate that 2 reproducible reliable routes exist that introduce genes of commercial interest into filamentous fungi.Deoxyribonucleic acid (DNA) constructs were also made in order to genetically programme fungi to express genes of commercial interest. Initial investigations focussed on the expression and secretion of human interleukin-6 (hIL-6). Such studies resulted in the production of this substance by filamentous fungi.DEVELOPMENT OF A TRANSFORMATION AND EXPRESSION SYSTEM IN ASPERGILLUS ORYZAE AND PENICILLIUM CHRYSOGENUM FOR CLONING THE NIA A GENE FOR NITRATE ASSIMILATION. IN PARTICULAR : 1. THE EQUIVALENT NIA A GENE FROM THE INDUSTRIAL STRAIN ASPERGILLUS ORYZAE WILL BE CLONED BY CROOS-HYBRIDISATION WITH THE AVAILABLE GENE FROM THE MODEL SPECIES ASPERGILLUS NIDULANS. 2. ATTEMPS WILL BE MADE TO INCREASE THE FREQUENCY OF TRANSFORMATION OF ASPERGILLUS ORYZAE BY VARYING THE PERIOD OF CELL WALL DIGESTION ON OTHER PARAMETERS AND BY INSERTION OF CERTAIN DNA SEQUENCES SUCH AS ANS 1 FROM ASPERGILLUS NIDULANS OR SEQUENCES FROM PUC 8. 3. THE CLONED GENE STRUCTURE WILL BE CHARACTERIZED BY S1 MAPPING AND BY NUCLEOTIDE SEQUENCING. THE POSITION AND STRUCTURE OF PROMOTOR REGION WILL BE DETERMINED. Champ scientifique natural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesbiological sciencesmicrobiologymycologymedical and health sciencesbasic medicinepharmacology and pharmacypharmaceutical drugsantibioticsnatural sciencesbiological sciencesgeneticsnucleotides Programme(s) FP1-BAP - Multiannual research action programme (EEC) in the field of biotechnology (BAP), 1985-1989 Thème(s) Data not available Appel à propositions Data not available Régime de financement CSC - Cost-sharing contracts Coordinateur University of St Andrews Contribution de l’UE Aucune donnée Adresse Irvine Building North Street KY16 9AL St Andrews Royaume-Uni Voir sur la carte Coût total Aucune donnée Participants (2) Trier par ordre alphabétique Trier par contribution de l’UE Tout développer Tout réduire LABORATORY OF MOLECULAR BIOLOGY Belgique Contribution de l’UE Aucune donnée Adresse GENT Voir sur la carte Coût total Aucune donnée TNO Pays-Bas Contribution de l’UE Aucune donnée Adresse RIJSWIJK Voir sur la carte Coût total Aucune donnée