Abstract
Marker chromosome are small supernumerary chromosomes that are sometimes associated with developmental abnormalities. Hence, the genes involved in such cases provide an interesting approach to understanding developmental abnormalities in man. As a first step towards isolating such sequences, marker chromosomes need complete characterization. By combining chromosome isolation by flow sorting and the “degenerate oligonucleotide primed — polymerase chain reaction”, we have constructed a DNA library specific for a marker chromosome found in a child with severe developmental abnormalities. We used fluorescent in situ hybridization of the library onto normal metaphase spreads (“reverse chromosome painting“) and were thus able to determine that the marker consists of the centromeric part of chromosome 7, the telomeric region of the long arm of chromosome 5 and the telomeric region of the short arm of the X-chromosome. Subsequently, we hybridized normal chromosome-specific libraries of the relevant chromosomes onto metaphases containing the marker chromosome (“forward chromosome painting”) and could in this manner establish the precise location of the different chromosome regions on the marker chromosome itself. This is a general approach suitable for outlining marker chromosomes in detail, and will aid the identification of the genes involved.
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References
Blennow E, Bröndum Nielsen K (1991) Molecular identification of a small supernumerary chromosome by in situ hybridization: diagnosis of an isochromosome 18p with probe L1. 84. Clin Genet 39:429–433
Buckton KE, Spowart G, Newton MS, Evans HJ (1985) Forty four probands with additional “marker” chromosome. Hum Genet 69:353–370
Callen DF, Freemantle CJ, Ringenbergs ML, Baker E, Eyre HJ, Romain D, Haan EA (1990) The isochromosome 18p syndrome: confirmation of cytogenetic diagnosis in nine cases by in situ hybridization. Am J Hum Genet 47:493–498
Callen DF, Eyre HJ, Ringenbergs ML, Freemantle CJ, Woodroffe P, Haan EA (1991) Chromosomal origin of small ring marker chromosomes in man: characterization by molecular genetics. Am J Hum Genet 48:769–782
Carter NP, Ferguson-Smith ME, Affara NA, Briggs H, Ferguson-Smith MA (1990) Study of X chromosome abnormality in XX males using bivariate flow karyotpye analysis and flow sorted dot blots. Cytometry 11:202–207
Carter NP, Ferguson-Smith MA, Perryman MT, Telenius H, Pelmear AH, Leversha MA, Glancy MT, Wood SL, Cook K, Dyson HM, Ferguson-Smith ME, Willat LR (1992) Reverse chromosome painting: a method for the rapid analysis of aberrant chromosomes in clinical cytogenetics. J Med Genet 29:299–307
Deng H-X, Yoshiura K, Dirks RW, Harada N, Hirota T, Tsukamoto K, Jinno Y, Niikawa N (1992) Chromosome-band-specific painting: chromosome in situ suppression hybridization using PCR products from a microdissected chromosome band as a probe pool. Hum Genet 89:13–17
Ehrenborg E, Larsson C, Stern I, Janson M, Powell DR, Luthman H (1992) Contiguous localization of the genes encoding human insulin-like growth factor binding proteins 1(IGBP1) and 3(IGBP3) on chromosome 7. Genomics 12:497–502
Friedrich U, Nielsen J (1974) Bisatellited extra small metacentric chromosome in newborns. Clin Genet 29:23–31
Jauch A, Daumer C, Lichter P, Murken J, Schroeder-Kurth T, Cremer T (1990) Chromosomal in situ suppression hybridization of human gonosomes and autosomes and its use in clinical cytogenetics. Hum Genet 85:145–150
Lichter P, Cremer T, Borden J, Manuelidis L, Ward DC (1988) Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries. Hum Genet 80:224–234
McDermid HE, Duncan AMV, Brasch KR, Holden JJA, Magenis E, Sheehy R, Burn J, Kardon N, Noel B, Schinzel A, Teshima I, White BN (1986) Characterization of the supernumerary chromosome in cat eye syndrome. Science 232:646–648
Meltzer PS, Guan X-Y, Burgess A, Trent JM (1992) Rapid generation of region specific probes by chromosome microdissection and their application. Nature Genet 1:24–28
Nielsen J, Rasmussen K (1975) Extra marker chromosome in newborn children. Hereditas 81:221–224
Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high sensitivity fluorescence hybridization. Proc Natl Acad Sci USA 83:2934–2938
Sachs ES, Hemel JO van, Hollander JC den, Jahoda MGJ (1987) Marker chromosomes in a series of 10000 prenatal diagnosis. Cytogenetic and follow up studies. Prenat Diagn 7:81–89
Schinzel A (1991) Tetrasomy 12p (Pallister-Killian syndrome). J Med Genet 28:122–125
Sillar R, Young BD (1981) A new method for the preparation of metaphases for flow analysis. J Histochem Cytochem 29P:74–78
Telenius H, Pelmear AH, Tunnacliffe A, Carter NP, Behmel A, Ferguson-Smith MA, Nordenskjöld M, Pfragner R, Ponder BAJ (1992) Cytogenetic analysis by chromosome painting using DOP-PCR amplified flow-sorted chromosomes. Genes Chrom Cancer 4:257–263
Trask BJ (1991) Fluorescence in situ hybridization: applications in cytogenetics and gene mapping. Trends Genet 7:149–154
Wahlström J, Borsgård J, Sabel K-G (1976) A case of trisomy 20? Clin Genet 9:187–191
Waltzer S, Breau G, Gerald PS (1969) A chromosome survey of 2400 normal newborn infants. J Pediatr 74:438–448
Warburton D (1984) Outcome of cases of de novo structural rearrangements diagnosed at amniocentesis. Prenat Diagn 4:69–80
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Blennow, E., Telenius, H., Larsson, C. et al. Complete characterization of a large marker chromosome by reverse and forward chromosome painting. Hum Genet 90, 371–374 (1992). https://doi.org/10.1007/BF00220461
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DOI: https://doi.org/10.1007/BF00220461