Abstract
To construct a mammalian artificial chromosome (MAC), telomere repeats and selectable markers were introduced into a 100 kb yeast artificial chromosome (YAC) containing human centromeric DNA. This YAC, which has a regular repeat structure of alpha-satellite DNA and centromere protein B (CENP-B) boxes, efficiently formed MACs that segregated accurately and bound CENP-B, CENP-C, and CENP-E. The MACs appear to be about 1-5 Mb in size and contain YAC multimers. Structural analyses suggest that the MACs have not acquired host sequences and were formed by a de novo mechanism. The accurate segregation of the MACs suggests they have potential as vectors for introducing genes into mammals.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Autoantigens*
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Cell Line
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Cell Line, Transformed
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Centromere / genetics
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Centromere Protein B
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Chromosomal Proteins, Non-Histone / chemistry
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Chromosomal Proteins, Non-Histone / genetics
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Chromosomes / genetics*
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Chromosomes, Artificial, Yeast / genetics*
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Chromosomes, Human, Pair 21 / genetics
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Cloning, Molecular
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DNA, Satellite / chemistry
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DNA, Satellite / genetics
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DNA-Binding Proteins*
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Genetic Vectors / genetics
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Humans
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In Situ Hybridization, Fluorescence
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Kinetochores
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Polymerase Chain Reaction
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Telomere / genetics
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Transfection
Substances
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Autoantigens
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CENPB protein, human
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Centromere Protein B
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Chromosomal Proteins, Non-Histone
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DNA, Satellite
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DNA-Binding Proteins
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centromere protein C
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centromere protein E