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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

Institut für Humangenetik u. Anthropologie, Universität Heidelberg, Im Neuenheimer Feld 328, 6900 Heidelberg, Germany [C. L., M. R. S., M. T., D. K., T. C.]; The Wistar Institute, Philadelphia, Pennsylvania 19104 [H. C. R.]; Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110 [E. D. G., M. V. O.]; Innere Klinik und Poliklinik (Tumorforschung), Universitätsklinikum Essen, Hufelandstrasse 55, 4300 Essen 1, Germany [R. B.]

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.

¹ This work was supported by grants from the Deutsche Forschungsgemeinschaft (Cr 59/10-1) and the Deutsche Krebshilfe (W23/90/Cr1). C. L. has been the recipient of a scholarship from the Konrad-Adenauer-Stiftung. H. R. was supported by an NIH postdoctoral fellowship (DHHS 1 F32 G11-12884). E. D. G. is a Lucille P. Markey Scholar, and his work was supported in part by a grant from the Lucille P. Markey Charitable Trust.

² To whom requests for reprints should be addressed.

Received 11/ 5/91. Accepted 2/26/92.

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