Premium
Search for the FSHD gene using cDNA selection in a region spanning 100 kb on chromosome 4q35
Author(s) -
van Deutekom Judith C. T.,
Hofker Marten H.,
Romberg Silke,
van Geel Michel,
Rommens Johanna,
Wright Tracy J.,
Hewitt Jane E.,
Padberg George W.,
Wijmenga Cisca,
Frants Rune R.
Publication year - 1995
Publication title -
muscle and nerve
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.025
H-Index - 145
eISSN - 1097-4598
pISSN - 0148-639X
DOI - 10.1002/mus.880181306
Subject(s) - genetics , selection (genetic algorithm) , gene , complementary dna , chromosome , biology , computational biology , computer science , artificial intelligence
Facioscapulohumeral muscular dystrophy (FSHD) is caused by deletions of 3.3‐kb tandemly repeated units contained within a large polymorphic Eco RI fragment close to the telomere of chromosome 4q. Since the rearrangements were assumed to interfere with the structure or function of the putative FSHD gene, the gene search was focused on cosmids containing these repeat units and, in addition, cosmids spanning 75 kb of upstream sequences. cDNA selection hybridization was applied to four overlapping cosmid clones, yielding a total of 150 putative cDNA clones. These clones showed a random distribution across the cosmid contig, except for three regions which contained a much larger number of clones. Nine cDNA clones hybridized to a 2.2‐kb Eco RI fragment, located 22 kb centromeric to the 3.3‐kb repeated units. This 2.2‐kb fragment showed evolutionary conservation, and analysis of the sequence by “GRAIL” predicted the presence of several exons. Transcripts homologous to this fragment could be identified but none of them originated from the 4q35 locus. Strikingly, most clones revealed 4–10 homologous loci, and no single copy clones could be isolated. These findings are in line with earlier observations by fluorescent in situ hybridization (FISH) showing hybridization of individual cosmid clones to multiple chromosomes. The presence of homologous regions on other chromosomes seriously complicates the cloning of the FSHD gene. © 1995 John Wiley & Sons, Inc.