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Detection of myc translocations in lymphoma cells by fluorescence in situ
hybridization with yeast artificial chromosomes
ML Veronese, M Ohta, J Finan, PC Nowell and CM Croce
Jefferson Cancer Institute, Jefferson Medical College, Philadelphia, PA
19107, USA.
Translocations involving chromosome 8 at band q24 and one of the Ig loci on
chromosomes 14q32, 22q11, and 2p11 are the hallmark of Burkitt's lymphoma
(BL). It has been previously observed that the exact localization of the
breakpoints at chromosome 8q24 can vary significantly from patient to
patient, scattering over a distance of more than 300 kb upstream of c-myc
and about 300 kb downstream of c- myc. To generate probes for fluorescence
in situ hybridization (FISH) that detect most c-myc translocations, we
screened a yeast artificial chromosome (YAC) library from normal human
lymphocytes by colony hybridization, using three markers surrounding the
c-myc gene as probes. We obtained 10 YAC clones ranging in size between 500
and 200 kb. Two nonchimeric clones were used for FISH on several BL cell
lines and patient samples with different breakpoints at 8q24. Our results
show that the YAC clones detected translocations scattered along
approximately 200 kb in both metaphase chromosomes and interphase nuclei.
The sensitivity, rapidity, and feasibility in nondividing cells render FISH
an important diagnostic tool. Furthermore, the use of large DNA fragments
such as YACs greatly simplifies the detection of translocations with widely
scattered breakpoints such as these seen in BL.
Volume 85,
Issue 8,
pp. 2132-2138,
04/15/1995
Copyright © 1995 by The American Society of Hematology
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