DEVELOPMENT OF A SEQUENCE-TAGGED SITE FOR THE CENTROMERE OF CHROMOSOME-10 - ITS USE IN CYTOGENETIC AND PHYSICAL MAPPING

We sequenced the alphoid centromere probe palpha10RP8 (D10Z1), aligned it to three published consensus sequences, and developed a sequence-t agged site (STS), sJRH-2, based upon oligonucleotide primers having tw o 3' mismatches with these consensus sequences. Polymerase chain react ion (PCR) amplification using genomic DNA from a somatic cell hybrid p anel representing all human chromosomes demonstrated amplification fro m only those cell lines containing chromosome 10. Fluorescence in situ hybridization of the amplified product demonstrated intense and speci fic 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 on e chimeric clone (consisting of DNA segments derived from chromosomes 5p and 10cen) confirmed specificity of the STS for the centromere of c hromosome 10. sJRH-2 provides a convenient cytogenetic marker for chro mosome 10, which will also be useful for physical mapping of the peric entromeric region of chromosome 10, a region that harbors the gene(s) for three forms of multiple endocrine neoplasia (types 2A, 2B, and fam ilial medullary thyroid carcinoma). The GenBank accession number for t he palpha10RP8 sequence is X63622.