Rapid generation of long synthetic tandem repeats and its application for analysis in human artificial chromosome formation.

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

SUBMITTER: Ebersole T 

PROVIDER: S-EPMC1197135 | biostudies-literature | 2005 Sep

REPOSITORIES: biostudies-literature