Abstract:
:Human artificial chromosomes (HACs) provide a unique opportunity to study kinetochore formation and to develop a new generation of vectors with potential in gene therapy. An investigation into the structural and the functional relationship in centromeric tandem repeats in HACs requires the ability to manipulate repeat substructure efficiently. We describe here a new method to rapidly amplify human alphoid tandem repeats of a few hundred base pairs into long DNA arrays up to 120 kb. The method includes rolling-circle amplification (RCA) of repeats in vitro and assembly of the RCA products by in vivo recombination in yeast. The synthetic arrays are competent in HAC formation when transformed into human cells. As short multimers can be easily modified before amplification, this new technique can identify repeat monomer regions critical for kinetochore seeding. The method may have more general application in elucidating the role of other tandem repeats in chromosome organization and dynamics.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Ebersole T,Okamoto Y,Noskov VN,Kouprina N,Kim JH,Leem SH,Barrett JC,Masumoto H,Larionov Vdoi
10.1093/nar/gni129keywords:
subject
Has Abstractpub_date
2005-09-01 00:00:00pages
e130issue
15eissn
0305-1048issn
1362-4962pii
33/15/e130journal_volume
33pub_type
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