Centromeric DNA of Kluyveromyces lactis

• Published in: Current genetics Vol. 18; no. 6; pp. 517 - 522
• Main Authors: Heus, J.J, Zonneveld, B.J.M, Steensma, H.Y, Berg, J.A. van den
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 01-12-1990; Berlin
• Subjects: Biological and medical sciences; Centromere; centromeres; Chromatin. Chromosome; Chromosome Mapping; Chromosomes, Fungal; Cloning, Molecular; DNA; DNA, Fungal - isolation & purification; Fundamental and applied biological sciences. Psychology; Genes, Fungal; Genomic Library; isolation; Kluyveromyces - genetics; Kluyveromyces lactis; Kluyveromyces marxianus var. lactis; Mitosis; Molecular and cellular biology; Molecular genetics; Plasmids; Restriction Mapping; selection methods; Fungi; Kluyveromyces marxianus var. lactis; Plasmid; Heterologous system; Stability; Centromere; Chromosome; Chromosome DNA; Ascomycetes; Molecular cloning; Thallophyta
• ISSN: 0172-8083; 1432-0983
• DOI: 10.1007/BF00327022

A direct selection method was used to isolate centromeres from a genomic library of the yeast Kluyveromyces lactis. The method is based on the lethality at high copy number of the ochre-suppressing tRNA gene SUP11. Five different chromosomal fragments were found that confer mitotic stability to plasmids containing a replication origin of K. lactis (KARS). In addition, KARS plasmids containing these fragments have a copy number of approximately one, and each of the five fragments hybridizes to a different chromosome of K. lactis. From these results we conclude that five of the six centromeres of K. lactis have been isolated. These centromeres do not function in S. cerevisiae.