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Long‐range chromatin analysis of the human MYC locus by pulsed‐field gel electrophoresis
Author(s) -
Mautner Josef,
Bornkamm Georg W.,
Polack Axel
Publication year - 1996
Publication title -
genes, chromosomes and cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.754
H-Index - 119
eISSN - 1098-2264
pISSN - 1045-2257
DOI - 10.1002/(sici)1098-2264(199608)16:4<247::aid-gcc4>3.0.co;2-y
Subject(s) - pulsed field gel electrophoresis , chromatin , gel electrophoresis , locus (genetics) , range (aeronautics) , biology , microbiology and biotechnology , genetics , materials science , dna , gene , genotype , composite material
The identification of cis‐acting regulatory elements has been greatly facilitated by the perception that nonnucleosomal regions of chromatin, including sites where transacting factors are bound, are hypersensitive to cleavage by nucleases. Hence, mapping of DNasel‐hypersensitive sites (HSs) has become particularly valuable for the detection of transcriptional control elements. The utility of this technique, however, may be constrained by the huge size of some eukaryotic gene domains or by the nonavailability of genomic probes. Apparently, both of these drawbacks hold true for the human protooncogene MYC . To overcome these limitations, we investigated the feasibility of mapping DNasel‐HSs in large restriction fragments. By using MYC ‐amplified cell lines, we devised a simple protocol that allowed for the detection of DNasel‐HSs at a distance of several hundred kb. In an attempt to identify additional regulatory elements required for MYC expression, we used this method to establish the long‐range chromatic structure of four MYC amplicons. This method has potential benefits and applications. Genes Chromosom Cancer 16:247–253 (1996) . © 1996 Wiley‐Liss, Inc.