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Isolation of a yeast artificial chromosome spanning the 8; 21 translocation breakpoint t(8; 21)(q22; q22. 3) in acute myelogenous leukemia

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America; (United States)
OSTI ID:5934594
;  [1]; ;  [2]; ; ;  [3];  [4]
  1. Univ. of Colorado Health Sciences Center, Denver (United States)
  2. Eleanor Roosevelt Inst. for Cancer Research, Denver, CO (United States)
  3. Univ. of Chicago, IL (United States)
  4. Univ. of Colorado Health Sciences Center, Denver (United States) Eleanor Roosevelt Inst. for Cancer Research, Denver, CO (United States)

The 8;21 translocation is one of the most common specific rearrangements in acute myelogenous leukemia. The authors have identified markers (D21S65 and a Not I boundary clone, Not-42, referred to as probe B) flanking the chromosome 21 translocation breakpoint (21q22.3) that demonstrate physical linkage in normal genomic DNA, by using at least three restriction endonucleases (Not I, Sac II, and BssHII), and that are located not more than 250-280 kilobases apart. Pulsed-field gel analysis of DNA from somatic cell hybrids containing the 8;21 translocation chromosomes demonstrates rearrangement of these markers. A 470-kilobase yeast artificial chromosome, YAC-Not-42, has been isolated that contains both probes. Mapping of {lambda} subclones constructed from YAC-Not-42 suggests that >95% of the yeast artificial chromosome DNA is located on the proximal (D21S65) side of the breakpoint. In situ hybridization studies using metaphase chromosomes from five acute myelogenous leukemia patients with the 8;21 translocation confirmed these results and demonstrated the translocation of probe B to the derivative chromosome 8. A chromosome walk of {approx} 39 kilobases from probe B has allowed identification of the breakpoint in DNA from a somatic cell hybrid containing the derivative chromosome 8. Since probe B contains conserved DNA sequences and is in close proximity to the translocation breakpoint, it may represent a portion of the involved gene on chromosome 21.

DOE Contract Number:
FG02-86ER60408
OSTI ID:
5934594
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 88:11; ISSN 0027-8424; ISSN PNASA
Country of Publication:
United States
Language:
English