Development of a sequence-tagged site for the centromere of chromosome 10: its use in cytogenetic and physical mapping |
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Authors: | James R Howe Terry C Lairmore Rosalie Veile Shenshen Dou Samuel A Wells Jr Helen Donis-Keller |
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Institution: | (1) Department of Surgery, Washington University School of Medicine, 63110 St. Louis, MO, USA;(2) Division of Human Molecular Genetics, Department of Surgery, Washington University School of Medicine, Box 8109, 63110 St. Louis, MO, USA |
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Abstract: | We sequenced the alphoid centromere probe p 10RP8 (D10Z1), aligned it to three published consensus sequences, and developed a sequence-tagged site (STS), sJRH-2, based upon oligonucleotide primers having two 3 mismatches with these consensus sequences. Polymerase chain reaction (PCR) amplification using genomic DNA from a somatic cell hybrid panel representing all human chromosomes demonstrated amplification from only those cell lines containing chromosome 10. Fluorescence in situ hybridization of the amplified product demonstrated intense and specific hybridization of the PCR product to 10p11.1-q11.1. A human genomic yeast artificial chromosome (YAC) library was screened using the sJRH-2 PCR assay, and five clones were identified. Sequence analysis of one chimeric clone (consisting of DNA segments derived from chromosomes 5p and 10cen) confirmed specificity of the STS for the centromere of chromosome 10. sJRH-2 provides a convenient cytogenetic marker for chromosome 10, which will also be useful for physical mapping of the pericentromeric region of chromosome 10, a region that harbors the gene(s) for three forms of multiple endocrine neoplasia (types 2A, 2B, and familial medullary thyroid carcinoma). The GenBank accession number for the p 10RP8 sequence is X63622. |
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