© 1994 Oxford University Press
RESEARCH-ARTICLE |
De novo formation of several features of a centromere following introduction of a Y alphoid YAC into mammalian cells
CRC Chromosome Molecular Biology Group, Department of Biochemistry, University of Oxford South Parks Road, Oxford OX1 3QU 1Department of Plant Sciences, University of Oxford South Parks Road, Oxford OX1 3RB, UK
*To whom correspondence should be addressed
Received January 20, 1994; Revised March 22, 1994; Accepted March 22, 1994
The DNA sequence requirements for mammalian centromere function have been Investigated by re-Introducing human YAC clones containing either centromeric or non-centromeric sequences Into hamster and human cells. All YACs integrated into the host chromosomes. In most cell lines produced by spheroplast fusion into hamster cells, intact copies of the YAC and a large amount of yeast DNA were found. Cell lines produced by lipofection Into human cells usually contained simple structures without yeast DNA. YACs containing Y alphoid DNA reformed several of the properties of a centromere, Including a cytogenetlcally visible constriction, CREST antiserum binding and disruption of anaphase chromosome movement. In contrast, YACs containing non-centromeric sequences produced none of these results. This work suggests that a few hundred kb of alphoid DNA is sufficient to reconstitute several Important features of a centromere.
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