Isolation of expressed sequences encoded by the human Xq terminal portion using microclone probes generated by laser microdissection.
The genes that cause a variety of neurologic and neuromuscular disorders have been mapped to the distal region of Xq. In an effort to isolate genes from this area, a regional genomic library of the distal 30% of Xq was constructed from a single metaphase spread by means of laser microdissection and single unique primer-polymerase chain reaction. Using pooled probes of 1000 clones from the genomic library, human brain cDNA libraries were screened for expressed sequences encoded by this region. From the 250,000 cDNA clones screened so far, 10 nonoverlapping sequences that mapped back to the target portion were isolated. The complete nucleotide sequences of these cDNA clones have been determined. Analysis of the sequences indicates that none has significant similarity to previously characterized primate genes. One sequence mapping to Xq27.3-qter contained an open reading frame of 281 amino acids and was expressed in every tissue tested. This gene, as well as others isolated in this manner, may prove to be a candidate gene for heritable disorders mapping to this region.
Ikeda Genosphere Project, ERATO, JRDC, University of Ottawa Faculty of Medicine, Ontario, Canada.
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Yokoi
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Hadano
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Kogi
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Kang
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Wakasa
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Ikeda
D000595:Amino Acid Sequence; D000782:Aneuploidy; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D002871:Chromosome Banding; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017931:DNA Primers; D005260:Female; D005333:Fetus; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D007621:Karyotyping; D008297:Male; D008969:Molecular Sequence Data; D009422:Nervous System Diseases / Q000235:genetics; D009468:Neuromuscular Diseases / Q000235:genetics; D015345:Oligonucleotide Probes; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods; D014960:X Chromosome
D017931:DNA Primers; D015345:Oligonucleotide Probes