title: 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.
authors.affiliation: The Murdoch Children's Research Institute, Royal Children's Hospital, Flemington Road, Melbourne 3052, Australia.
authors.forename: R
authors.initials: R
authors.lastname: Saffery
authors.forename: L H
authors.initials: LH
authors.lastname: Wong
authors.forename: D V
authors.initials: DV
authors.lastname: Irvine
authors.forename: M A
authors.initials: MA
authors.lastname: Bateman
authors.forename: B
authors.initials: B
authors.lastname: Griffiths
authors.forename: S M
authors.initials: SM
authors.lastname: Cutts
authors.forename: M R
authors.initials: MR
authors.lastname: Cancilla
authors.forename: A C
authors.initials: AC
authors.lastname: Cendron
authors.forename: A J
authors.initials: AJ
authors.lastname: Stafford
authors.forename: K H
authors.initials: KH
authors.lastname: Choo
mesh_terms: D002460:Cell Line; D002503:Centromere; D002877:Chromosomes, Human; D018014:Gene Transfer Techniques; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016615:Telomere