Objective CONSTRUCTION OF EFFICIENT CLONING VECTORS ABLE TO TRANSFORM KLUYVEROMYCES AND OTHER YEASTS USED IN THE DAIRY INDUSTRY. The genus Kluyveromyces is important in dairy industries and lately the pKD1 plasmid as well as the KARS based vectors have proved to be promising instruments in the physiological studies and genetic engineering of dairy yeasts.Efficient expression vectors have been developed for the yeasts of industrial importance, in particular, the dairy yeasts.Research has involved:structural and functional characterization of the plasmid pKD1 and its derivative vectors;analysis of the mechanism that gives autonomy and stability to the vectors derived from 2 and pKD1;study of physiological regulation in Kluyveromyces lactis as a production organism.The molecular structures of the plasmid pKD1 and of the autonomously replicating KARS sequence from K lactis chromosome were established. Plasmid and host functions important for the stable maintenance of the yeast plasmid were examined. Highly stable forms of pKD1 derived vectors were obtained. The host range of pKD1 was extended to several other species of Kluyveromyces. Shuttle deoxyribonucleic acid (DNA) banks were constructed for Kluyveromyces and Saccharomyces species. By combining the K lactis killer toxin gene sequence and pKD1 derived vectors, performing secretion vectors were obtained. Human interleukin-1 beta could be produced at high levels. To better control the gene expression of K lactis, several promoters were studied; in particular, new knowledge was obtained on the regulation of lactose metabolism and in the alcohol dehydrogenase system.CONSTRUCTION OF NEW CLONING VECTORS FOR THE YEAST KLUYVEROMYCES. STEPS : 1- DESCRIPTION AND SEQUENCING OF THREE NEW PLASMIDS RECENTLY DISCOVERED IN KLUYVEROMYCES SPECIES. 2- ISOLATION OF AUXOTROPHIC MUTATIONS IN KLUYVEROMYCES. 3- IDENTIFICATION OF REPLICATION ORIGINS ON THE NEW PLASMIDS. 4- IMPROVEMENT OF TRANSFORMATION EFFICIENCY USING CONSTRUCTIONS CONTAINING ORIGINS OF REPLICATIONS FROM THE NEW PLASMIDS AND THE S.CEREVISIAE MARKER URA3. Fields of science natural sciencesbiological sciencesgeneticsDNAagricultural sciencesanimal and dairy sciencedairymedical and health sciencesmedical biotechnologygenetic engineeringnatural scienceschemical sciencesorganic chemistryalcoholsnatural sciencesbiological sciencesgeneticschromosomes Programme(s) FP1-BAP - Multiannual research action programme (EEC) in the field of biotechnology (BAP), 1985-1989 Topic(s) Data not available Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator Institut Curie Address Université de paris xi (université paris-sud) 15 rue georges clémenceau 91405 Orsay France See on map EU contribution € 0,00 Participants (2) Sort alphabetically Sort by EU Contribution Expand all Collapse all INSTITUT FÜR MIKROBIOLOGIE Germany EU contribution € 0,00 Address Düsseldorf See on map Università degli Studi di Roma Italy EU contribution € 0,00 Address Roma See on map
