Isolation of a yeast artificial chromosome spanning the 8; 21 translocation breakpoint t(8; 21)(q22; q22. 3) in acute myelogenous leukemia
- Univ. of Colorado Health Sciences Center, Denver (United States)
- Eleanor Roosevelt Inst. for Cancer Research, Denver, CO (United States)
- Univ. of Chicago, IL (United States)
- 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
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
BIOLOGICAL MARKERS
CHROMOSOMES
DISEASES
DNA HYBRIDIZATION
DNA SEQUENCING
EUMYCOTA
FUNGI
HEREDITARY DISEASES
HETEROCHROMOSOMES
HUMAN CHROMOSOME 21
HUMAN CHROMOSOMES
HYBRIDIZATION
IMMUNE SYSTEM DISEASES
LEUKEMIA
MICROORGANISMS
MYELOID LEUKEMIA
NEOPLASMS
PATIENTS
PLANTS
STRUCTURAL CHEMICAL ANALYSIS
TRANSLOCATION
YEASTS