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

  1. G Hadlaczky,
  2. T Praznovszky,
  3. I Cserpán,
  4. J Keresö,
  5. M Péterfy,
  6. I Kelemen,
  7. E Atalay,
  8. A Szeles,
  9. J Szelei, and
  10. V Tubak
  1. Institute of Genetics Biological Research Center, Hungarian Academy of Sciences, Szeged.

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.

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  1. PNAS September 15, 1991 vol. 88 no. 18 8106-8110
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