Gene mapping and physical arrangements of human chromatin in transformed, hybrid cells: fluorescent and autoradiographic in situ hybridization compared

Abstract

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.