Centromere formation in mouse cells cotransformed with human DNA and a dominant marker gene.
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.
Hadlaczky
G
G
Institute of Genetics Biological Research Center, Hungarian Academy of Sciences, Szeged.
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
D000818:Animals; D002503:Centromere / Q000648:ultrastructure*; D003001:Cloning, Molecular; D004247:DNA / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D019868:Kanamycin Kinase; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010770:Phosphotransferases / Q000235:genetics; D015183:Restriction Mapping
D004247:DNA; D010770:Phosphotransferases; D019868:Kanamycin Kinase