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A human supernumerary minichromosome (MC), previously identified as a derivative of chromosome 9, has been introduced into Chinese hamster ovary (CHO) cells by means of cell fusion. A hybrid clone containing the MC as the only free human chromosome was isolated. A selectable marker gene (neo) inserted into a yeast artificial chromosome (YAC) has been successfully targeted to the MC centromeric DNA via co-transfection with chromosome-9-specific alpha satellite DNA. In situ hybridization and Southern blotting experiments demonstrated that the intact neo gene was integrated into the MC centromeric DNA. Studies on the clonal distribution and on the stability of the MC either in the presence or in the absence of the selective agent have been carried out. The MC is susceptible to further manipulations and may thus represent a model for the construction of a large-capacity vector for somatic gene therapy.

Original publication

DOI

10.1089/hum.1996.7.9-1103

Type

Journal article

Journal

Human gene therapy

Publication Date

06/1996

Volume

7

Pages

1103 - 1109

Addresses

Departimento di Genetica e Microbiologia A Buzzati Traverso, Pavia, Italy.

Keywords

Cell Line, CHO Cells, Chromosomes, Artificial, Yeast, Chromosomes, Human, Pair 9, Centromere, Animals, Humans, DNA, Bacterial, DNA, Satellite, DNA Probes, Blotting, Southern, Gene Targeting, Transfection, Cricetinae