Abstract
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 α 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.
Overview summary
The bacterial neo gene as a selectable marker has been targeted to the centromeric DNA of a human accessory minichromosome (MC) via co-transfection with specific α satellite DNA sequences. The in vitro stability of the MC has been demonstrated. Experiments of size reduction by X-ray exposure and telomere-directed fragmentation are currently made in view of the potential function of the MC as a vector for somatic gene therapy.
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