Gene mapping and physical arrangements of human chromatin in transformed, hybrid cells: fluorescent and autoradiographic in situ hybridization compared.
We compare a fluorescent in situ hybridization technique, using N-acetoxy-2-acetylaminofluorene (N-ACO-AAF) modified DNA adducts, with 3H-labeled DNA in situ hybridization for visualizing human transgenomes in HRAS1-selected, chromosome-mediated gene transfer (CMGT), and mapping chromosomal SV40 in an SV40-transformed, human-mouse hybrid cell line. We demonstrate that individual HRAS1-CMGTs may contain multiple fragments of human chromatin. We deduce that the CMGT process can involve interstitial loss of mouse chromatin. We conclude that the N-ACO-AAF technique gives finer resolution than 3H-labeled in situ hybridization. However, 3H-labeling is more sensitive and has allowed us to sublocalize SV40 in C121 to the region 7q31-35.
D000818:Animals; D002472:Cell Transformation, Viral; D002843:Chromatin / Q000032:analysis / Q000235:genetics*; D002874:Chromosome Mapping; D002906:Chromosomes, Human, 6-12 and X; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes; D005456:Fluorescent Dyes; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D009693:Nucleic Acid Hybridization; D013539:Simian virus 40 / Q000235:genetics
D002843:Chromatin; D005456:Fluorescent Dyes; D004247:DNA; D004262:DNA Restriction Enzymes