A panel of irradiation-reduced hybrids selectively retaining human chromosome 11p13: Their structure and use to purify the WAGR gene complex

Thomas M Glaser, Elise Rose, Helvise Morse, David Housman, Carol Jones

Research output: Contribution to journalArticlepeer-review

45 Scopus citations


The irradiation-fusion technique offers a means to isolate intact subchromosomal fragments of one mammalian species in the genetic background of another. Irradiation-reduced somatic cell hybrids can be used to construct detailed genetic and physical maps of individual chromosome bands and to systematically clone genes responsible for hereditary diseases on the basis of their chromosomal position. To assess this strategy, we constructed a panel of hybrids that selectively retain the portion of human chromosome band 11p13 that includes genes responsible for Wilms tumor, aniridia, genitourinary anomalies, and mental retardation (constituting the WAGR syndrome). A hamster-human hybrid containing the short arm of chromosome 11 as its only human DNA (J1 - 11) was γ-irradiated and fused to a Chinese hamster cell line (CHO-K1). We selected secondary hybrid clones that express MIC1 but not MER2, cell-surface antigens encoded by bands 11p13 and 11p15, respectively. These clones were characterized cytogenetically by in situ hybridization with human repetitive DNA and were tested for their retention of 56 DNA, isozyme, and antigen markers whose order on chromosome 11p is known. These cell lines appear to carry single, coherent segments of 11p spanning MIC1, which range in size from 3000 kb to more than 50,000 kb and which are generally stable in the absence of selection. In addition to the selected region of 11p13, two cell lines carry extra fragments of the human centromere and two harbor small, unstable segments of 11p15. As a first step to determine the size and molecular organization of the WAGR gene complex, we analyzed a subset of reduced hybrids by pulsed-field gel electrophoresis. A small group of NotI restriction fragments comprising the WAGR complex was detected in Southern blots with a cloned Alu repetitive probe. One of the cell lines (GH3A) was found to carry a stable ∼3000-kb segment of 11p13 as its only human DNA. The segment encompasses MIC1, a recurrent translocation breakpoint in acute T-cell leukemia (TCL2), and most or all of the WAGR gene complex, but does not include the close flanking markers D11S16 and ΔJ. This hybrid forms an ideal source of molecular clones for the developmentally fascinating genes underlying the WAGR syndrome.

Original languageEnglish (US)
Pages (from-to)48-64
Number of pages17
Issue number1
StatePublished - 1990
Externally publishedYes

ASJC Scopus subject areas

  • Genetics


Dive into the research topics of 'A panel of irradiation-reduced hybrids selectively retaining human chromosome 11p13: Their structure and use to purify the WAGR gene complex'. Together they form a unique fingerprint.

Cite this