Ewing's tumor X mouse hybrids expressing the MIC2 antigen: analyses using fluorescence CDD-banding and non-isotopic ISH View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1992-01

AUTHORS

M. Dworzak, C. Stock, S. Strehl, H. Gadner, P. F. Ambros

ABSTRACT

We present a highly sensitive method that has been applied to map the chromosomal origin of the prominent cell surface antigen of Ewing's tumor cells recognized by monoclonal antibody HBA-71. The technique allows an unambiguous identification of human chromosomal material in interspecific cell hybrids. This is achieved by fluorescent in situ hybridization of biotinylated total human DNA, followed by high resolution fluorescence banding with the chromomycin/distamycin/DAPI triple stain. An advantage of this method is that all signals can be visualized in one single operation by simply switching the appropriate filter blocks. The protocol has proved extremely useful in gene mapping by means of interspecific cell hybrids, a technique that depends on the accurate and unambiguous recognition of the relevant (e.g., human) genetic material in the clonal genome. Our studies confirm that this antigen is the product of the MIC2 gene, which is so far the only well-studied pseudoautosomal gene in man, and which is located on the short arms of chromosomes X and Y. Furthermore, no influence of the Ewing's tumor-specific translocation t(11;22) on MIC2 expression could be discerned. More... »

PAGES

273-278

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00197258

DOI

http://dx.doi.org/10.1007/bf00197258

DIMENSIONS

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PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/1733829


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46 schema:description We present a highly sensitive method that has been applied to map the chromosomal origin of the prominent cell surface antigen of Ewing's tumor cells recognized by monoclonal antibody HBA-71. The technique allows an unambiguous identification of human chromosomal material in interspecific cell hybrids. This is achieved by fluorescent in situ hybridization of biotinylated total human DNA, followed by high resolution fluorescence banding with the chromomycin/distamycin/DAPI triple stain. An advantage of this method is that all signals can be visualized in one single operation by simply switching the appropriate filter blocks. The protocol has proved extremely useful in gene mapping by means of interspecific cell hybrids, a technique that depends on the accurate and unambiguous recognition of the relevant (e.g., human) genetic material in the clonal genome. Our studies confirm that this antigen is the product of the MIC2 gene, which is so far the only well-studied pseudoautosomal gene in man, and which is located on the short arms of chromosomes X and Y. Furthermore, no influence of the Ewing's tumor-specific translocation t(11;22) on MIC2 expression could be discerned.
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