De novo formation of several features of a centromere following introduction of a Y alphoid YAC into mammalian cells |
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Authors: | Larin Zoia; Fricker Mark D; Tyler-Smith Chris |
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Institution: | 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 |
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Abstract: | The DNA sequence requirements for mammalian centromere functionhave been Investigated by re-Introducing human YAC clones containingeither centromeric or non-centromeric sequences Into hamsterand human cells. All YACs integrated into the host chromosomes.In most cell lines produced by spheroplast fusion into hamstercells, intact copies of the YAC and a large amount of yeastDNA were found. Cell lines produced by lipofection Into humancells usually contained simple structures without yeast DNA.YACs containing Y alphoid DNA reformed several of the propertiesof a centromere, Including a cytogenetlcally visible constriction,CREST antiserum binding and disruption of anaphase chromosomemovement. In contrast, YACs containing non-centromeric sequencesproduced none of these results. This work suggests that a fewhundred kb of alphoid DNA is sufficient to reconstitute severalImportant features of a centromere. |
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