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Medizin - Open Access LMU - Teil 09/22 Podcast
Metaphase and Interphase Cytogenetics with Alu-PCR-amplified Yeast Artificial Chromosome Clones Containing the BCR Gene and the Protooncogenes c-raf-1, c-fms, and c-erbB-21

Metaphase and Interphase Cytogenetics with Alu-PCR-amplified Yeast Artificial Chromosome Clones Containing the BCR Gene and the Protooncogenes c-raf-1, c-fms, and c-erbB-21

Medizin - Open Access LMU - Teil 09/22 vor 34 Jahren

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A human yeast artificial chromosome (YAC) library was screened by
polymerase chain reaction with oligonucleotide primers defined for
DNA sequences of the BCR gene and the protooncogenes c-raf-1,
c-fms, and c-erB-2. Alu-PCR-generated human DNA sequences were
obtained from the respective YAC clones and used for fluorescence
in situ hybridization experiments under suppression conditions.
After chromosomal in situ suppression hybridization to GTG-banded
human prometaphase chromosomes, seven of nine initially isolated
YAC clones yielded strong signals exclusively in the chromosome
bands containing the respective genes. Two clones yielded
additional signals on other chromosomes and were excluded from
further tests. The band-specific YACs were successfully applied to
visualize specific structural chromosome aberrations in peripheral
blood cells from patients with myelodysplasia exhibiting
del(5)(q13q34), chronic myeloid leukemia and acute lymphocytic
leukemia with t(9;22)(q34;q11), acute promyelocytic leukemia (M3)
with t(15;17)(q22;q21), and in a cell line established from a
proband with the constitutional translocation t(3;8)(p14.2;q24). In
addition to the analysis of metaphase spreads, we demonstrate the
particular usefulness of these YAC clones in combination with whole
chromosome painting to analyze specific chromosome aberrations
directly in the interphase nucleus.