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Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation.


ABSTRACT: Neocentromeres (NCs) are fully functional centromeres that arise ectopically in noncentromeric regions lacking alpha-satellite DNA. Using telomere-associated chromosome truncation, we have produced a series of minichromosomes (MiCs) from a mardel(10) marker chromosome containing a previously characterized human NC. These MiCs range in size from approximately 0.7 to 1.8 Mb and contain single-copy intact genomic DNA from the 10q25 region. Two of these NC-based Mi-Cs (NC-MiCs) appear circular whereas one is linear. All demonstrate stability in both structure and mitotic transmission in the absence of drug selection. Presence of a functional NC is shown by binding a host of key centromere-associated proteins. These NC-MiCs provide direct evidence for mitotic segregation function of the NC DNA and represent examples of stable mammalian MiCs lacking centromeric repeats.

SUBMITTER: Saffery R 

PROVIDER: S-EPMC33277 | biostudies-literature | 2001 May

REPOSITORIES: biostudies-literature

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Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation.

Saffery R R   Wong L H LH   Irvine D V DV   Bateman M A MA   Griffiths B B   Cutts S M SM   Cancilla M R MR   Cendron A C AC   Stafford A J AJ   Choo K H KH  

Proceedings of the National Academy of Sciences of the United States of America 20010501 10


Neocentromeres (NCs) are fully functional centromeres that arise ectopically in noncentromeric regions lacking alpha-satellite DNA. Using telomere-associated chromosome truncation, we have produced a series of minichromosomes (MiCs) from a mardel(10) marker chromosome containing a previously characterized human NC. These MiCs range in size from approximately 0.7 to 1.8 Mb and contain single-copy intact genomic DNA from the 10q25 region. Two of these NC-based Mi-Cs (NC-MiCs) appear circular where  ...[more]

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