title: Cloning and sequencing of the URA3 gene of Kluyveromyces marxianus CBS 6556.
abstract: The URA3 gene, coding for orotidine-5'-phosphate decarboxylase, from Kluyveromyces marxianus CBS 6556, was isolated from a genomic DNA library. The K. marxianus URA3 gene encodes a protein of 267 amino acids with a calculated molecular weight of 29.3 kDa. Comparison of the K. marxianus protein with the corresponding enzymes of Saccharomyces cerevisiae and Kluyveromyces lactis showed amino acid sequence identities of 81% and 88%, respectively. Using contour-clamped homogenous electric field gel electrophoresis, the genomic copy was found to be located on chromosome VI. We have used the cloned gene for the construction of a K. marxianus leu2 mutant. This mutant contains no heterologous sequences, which is essential to make it acceptable for application in the food industry.
authors.affiliation: Laboratorium voor Biochemie en Moleculaire Biologie, Vrije Universiteit, Amsterdam, The Netherlands.
authors.forename: R J
authors.initials: RJ
authors.lastname: Bergkamp
authors.forename: R H
authors.initials: RH
authors.lastname: Geerse
authors.forename: J M
authors.initials: JM
authors.lastname: Verbakel
authors.forename: R J
authors.initials: RJ
authors.lastname: Planta
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular; D005800:Genes, Fungal / Q000235:genetics*; D007716:Kluyveromyces / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics
chemical_list: D009964:Orotidine-5'-Phosphate Decarboxylase