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Construction of a Yeast Artificial Chromosome Contig Spanning the Spinal Muscular Atrophy Disease Gene Region
Patrick W. Kleyn, Ching H. Wang, Lyndon L. Lien, Emilia Vitale, Jie Pan, Barbara M. Ross, Adina Grunn, David A. Palmer, Dorothy Warburton, Linda M. Brzustowicz, Louis M. Kunkel and T. Conrad Gilliam
Proceedings of the National Academy of Sciences of the United States of America
Vol. 90, No. 14 (Jul. 15, 1993), pp. 6801-6805
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/2362594
Page Count: 5
You can always find the topics here!Topics: DNA, Chromosomes, Polymerase chain reaction, Genomics, Libraries, Genes, Genetic loci, Nervous system diseases, Genetic diseases, Muscular diseases
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The childhood spinal muscular atrophies (SMAs) are the most common, serious neuromuscular disorders of childhood second to Duchenne muscular dystrophy. A single locus for these disorders has been mapped by recombination events to a region of 0.7 centimorgan (range, 0.1-2.1 centimorgans) between loci D5S435 and MAP1B on chromosome 5q11.2-13.3. By using PCR amplification to screen yeast artificial chromosome (YAC) DNA pools and the PCR-vectorette method to amplify YAC ends, a YAC contig was constructed across the disease gene region. Nine walk steps identified 32 YACs, including a minimum of seven overlapping YAC clones (average size, 460 kb) that span the SMA region. The contig is characterized by a collection of 30 YAC-end sequence tag sites together with seven genetic markers. The entire YAC contig spans a minimum of 3.2 Mb; the SMA locus is confined to roughly half of this region. Microsatellite markers generated along the YAC contig segregate with the SMA locus in all families where the flanking markers (D5S435 and MAP1B) recombine. Construction of a YAC contig across the disease gene region is an essential step in isolation of the SMA-encoding gene.
Proceedings of the National Academy of Sciences of the United States of America © 1993 National Academy of Sciences