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Centromere formation in mouse cells cotransformed with human DNA and a dominant marker gene.

ABSTRACT: A 13,863-base-pair (bp) putative centromeric DNA fragment has been isolated from a human genomic library by using a probe obtained from metaphase chromosomes of human colon carcinoma cells. The abundance of this DNA was estimated to be 16-32 copies per genome. Cotransfection of mouse cells with this sequence and a selectable marker gene (aminoglycoside 3'-phosphotransferase type II, APH-II) resulted in a transformed cell line carrying an additional centromere in a dicentric chromosome. This centromere was capable of binding an anti-centromere antibody. In situ hybridization demonstrated that the human DNA sequence as well as the APH-II gene and vector DNA sequences were located only in the additional centromere of the dicentric chromosome. The extra centromere separated from the dicentric chromosome, forming a stable minichromosome. This functional centromere linked to a dominant selectable marker may be a step toward the construction of an artificial mammalian chromosome.

SUBMITTER: Hadlaczky G 

PROVIDER: S-EPMC52455 | biostudies-other | 1991 Sep

REPOSITORIES: biostudies-other

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Centromere formation in mouse cells cotransformed with human DNA and a dominant marker gene.

Hadlaczky G G   Praznovszky T T   Cserpán I I   Keresö J J   Péterfy M M   Kelemen I I   Atalay E E   Szeles A A   Szelei J J   Tubak V V  

Proceedings of the National Academy of Sciences of the United States of America 19910901 18

A 13,863-base-pair (bp) putative centromeric DNA fragment has been isolated from a human genomic library by using a probe obtained from metaphase chromosomes of human colon carcinoma cells. The abundance of this DNA was estimated to be 16-32 copies per genome. Cotransfection of mouse cells with this sequence and a selectable marker gene (aminoglycoside 3'-phosphotransferase type II, APH-II) resulted in a transformed cell line carrying an additional centromere in a dicentric chromosome. This cent  ...[more]