Comparison of methods for identifying transcription units and transcription map of the Werner syndrome gene region.
To isolate a human disease gene by positional cloning, a critical step is the identification of candidate genes from a targeted genomic region. We used cDNA selection, exon trapping, and genomic sequencing to identify 12 transcription units from a 1.4-Mb genomic region containing the Werner syndrome gene (WRN). This included sequencing of 650 kb in the region of the WRN gene, to date, the most DNA sequenced as part of a positional cloning effort. The result of this combined method was significant overlap among the transcription units identified by each method; yet, no one method identified all of the transcription units. We present here a comparison of the effectiveness and efficiency of these methods and present a transcription map of the Werner syndrome gene region.
Hisama
F M
FM
Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Oshima
J
J
Yu
C E
CE
Fu
Y H
YH
Mulligan
J
J
Weissman
S M
SM
Schellenberg
G D
GD
D015152:Blotting, Northern; D002874:Chromosome Mapping / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods*; D020224:Expressed Sequence Tags; D006801:Humans; D009154:Mutation / Q000235:genetics; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic / Q000235:genetics*; D014898:Werner Syndrome / Q000235:genetics*