Abstract
Employing a modified Goss-Harris irradiation fusion protocol, we have generated a panel of somatic cell hybrids containing various overlapping fragments of the Xcen-Xp11.4 interval. This region of the human X chromosome is known to carry genes for several hereditary eye diseases including retinitis pigmentosa (RP2), congenital stationary night blindness (CSNB-1) and Norrie disease. These hybrid cell lines were employed to isolate 17 new DNA probes by making use of the Alu polymerase chain reaction (PCR) method and subsequent cloning of the PCR products in a plasmid vector. With these probes, we have characterized two previously described microdeletions spanning the Norrie locus; these deletions have enabled us to subdivide the Xp11.4-p11.3 region into three defined intervals.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benham F, Hart K, Crolla J, Bobrow M, Francavilla M, Goodfellow PN (1989) A method for generating hybrids containing nonselected fragments of human chromosomes. Genomics 4:509–517
Chapelle A de la, Sankila EM, Lindlöf N, Aula P, Norio R (1985) Norrie disease caused by a gene deletion allowing carrier detection and prenatal diagnosis. Clin Genet 28:317–320
Cotter FE, Hampton GM, Nasipuri S, Bodmer WF, Young BD (1990) Rapid isolation of human chromosome-specific DNA probes from a somatic cell hybrid. Genomics 7:257–263
Cremer T, Lichter P, Borden J, Waed DC, Manuelidis L (1988) Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosomespecific library probes. Hum Genet 80:235–246
Diergaarde PJ, Wieringa B, Bleeker-Wagemaker EM, Sims KB, Breakefield XO, Ropers H-H (1990) Physical fine mapping of a deletion spanning the Norrie gene. Hum Genet 84:22–26
Donnai D, Mountford RC, Read AP (1988) Norrie disease resulting from a gene deletion: clinical features and DNA studies. J Med Genet 25:73–78
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Feinberg AP, Vogelstein B (1984) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Addendun. Anal Biochem 137:266–267
Gal A, Bleeker-Wagemakers EM, Wienker TF, Warburg M, Ropers H-H (1985) Localization of the gene for Norrie disease by linkage to the DXS 7 locus (abstract). (8th International Workshop on Human Gene Mapping) Cytogenet Cell Genet 40:633
Gal A, Wieringa B, Smeets DF, Bleeker-Wagemakers L, Ropers H-H (1986) Submicroscopic interstitial deletion of the X chromosome explains a complex genetic syndrome dominated by Norrie disease. Cytogenet Cell Genet 42:219–224
Goss SJ, Harris H (1975) New method for mapping genes in human chromosomes. Nature 255:680–683
Lafrenière RG, Brown CJ, Powers VE, Carrel L, Davies KE, Barker DF, Willard HF (1991) Physical fine mapping of 60 DNA markers in the p21. 1–q21. 3 region of the human X chromosome. Genomics 11:352–363
Ledbetter SA, Nelson DL, Warren ST, Ledbetter DH (1990) Rapid isolation of DNA probes within specific chromosome regions by interspersed repetitive sequence polymerase chain reaction. Genomics 6:475–481
Lichter P, Cremer T, Borden J, 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
Mahtani MM, Lafrenière RG, Kruse TA, Willard HF (1991) An 18-locus linkage map of the pericentromeric region of the human X chromosome. Genomics 10:849–857
Miller SA, Dykes DD, Polesky HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16:1215
Musarella MA, Weleber RG, Murphey WH, Young RSL, AnsonCartwright L, Mets M, Kraft SP, Polemeno R, Litt M, Worton RG (1989) Assignment of the gene for complete X-linked congenital stationary night blindness (CSNB1) to Xp11. 3. Genomics 5:727–737
Nelson DL, Ledbetter SA, Corbo L, Victoria MF, Ramirez-Solis R, Webster TD, Ledbetter DH, Caskey CT (1989) Alu polymerase chain reaction: a method for rapid isolation of human specific sequences from complex DNA sources. Proc Natl Acad Sci USA 86:6686–6690
Ott J, Bhattacharya S, Chen JD, Denton MJ, Donald J, Dubay C, Farrar GJ, Fishman GA, Frey D, Gal A (1990) Localizing multiple X chromosome linked retinitis pigmentosa loci using multilocous homogeneity tests. Proc Natl Acad Sci USA 87:701–704
Sims KB, Chapelle A de la, Norio R, Sankila EM, Hsu Y-PP, Reinhart WB, Corey TJ, Ozelius L, Powell JF, Bruns G, Gusella JG, Murphy DL, Breakefield XO (1989) Monoamine oxidase deficiency in males with an X chromosome deletion. Neuron 2:1069–1076
Sinke RJ, Suijkerbuijk RF, Herbergs J, Janssen H, Cassiman JJ, Geurts van Kessel A (1992) Generation of a panel of somatic cell hybrids containing fragments of human chromosome 12p by X-ray irradiation and cell fusion. Genomics 12:206–213
Wieacker P, Davies KE, Cooke HJ, Pearson PL, Williamson PL, Bhattacharya S, Zimmer J, Ropers H-H (1984) Towards a comlete linkage map of the human X chromosome: regional assignment of 16 clones single copy DNA sequences employing a panel of somatic cell hybrids. Am J Hum Genet 36:265–270
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Berger, W., Meindl, A., de Leeuw, B. et al. Generation and characterization of radiation reduced cell hybrids and isolation of probes from the proximal short arm of the human X chromosome. Hum Genet 90, 243–246 (1992). https://doi.org/10.1007/BF00220070
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00220070