Centromeric DNA of Kluyveromyces lactis

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 plas...

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Bibliographic Details
Published in:Current genetics 1990-12, Vol.18 (6), p.517-522
Main Authors: Heus, J.J, Zonneveld, B.J.M, Steensma, H.Y, Berg, J.A. van den
Format: Article
Language:English
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
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Summary: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.
ISSN:0172-8083
1432-0983
DOI:10.1007/BF00327022