Human Molecular Genetics, Vol. 1, No. 6 451
Insertion/deletion polymorphism at D19S95 associated with the myotonic dystrophy CTG repeat
Taq\ and Bsu36\ polymorphisms in the human glycoprotein Iba gene (GPIBa:)
S.R.Crow*, H.G.Harley, J.D.Brook1, S.A.Rundle and D.J.Shaw Institute of Medical Genetics, University of Wales College of Medicine, Heath Park, Cardiff, CF4 6EJ, UK and 1 Centre for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
E.J.Petersen* and R.I.Handin Brigham and Women's Hospital, Department of Hematology and Harvard Medical School, Department of Medicine, Boston, MA, USA
Polymorphism: pBB0.7 identifies a 1.0 kb insertion/deletion RFLP with the enzymes EcoRV (10.5 kb, 9.5 kb), Eco9A (10 kb, 9 kb), HincJl (14 kb, 13 kb), PvuU (5.0 kb, 4.0 kb) and BgK (5.0 kb, 4.0 kb). Frequency: For the EcoRV RFLP, allele frequencies studied in 90 unrelated individuals are as follows: Al (10.5) 0.56 A2 (9.5) 0.44 Heterozygosity = 0.49.
Polymorphisms: Taql identifies a polymorphic site, detected as the presence or absence of a 4.0 kb fragment, with constant bands at 0.5, 0.7 and 1.1 kb. Bsu36I identifies a polymorphic site, detected as the presence or absence of a 6.9 kb fragment, with constant bands at 6.0 kb. Frequency: Taql studied in ten unrelated Caucasian individuals and Bsu36I in 15. Taql
+ 0.15 - 0.85 Heterozygosity = 0.26
Bsu36I
+ 0.20 - 0.80 Heterozygosity = 0.32
Not Polymorphic For: AccI, Apal, BamHI, Bell, BgU, Bgin, Not Polymorphic For: Aval, BamHl, Banl, BgH, HinfU, HindSR, Dral, EcoRl, EcoRV, HaeDI, HindHI, Hinfl, Hpafl, Kpnl, Ncol, Pstl, PvuE, Sad, Sau3AI, Xbal, XmnI, as studied in 10 unrelated HaeUl, Mspl, Pstl, Rsal, Sail, Smal, Sphl, Taql, Xbal. Caucasians. Chromosomal Localisation: pBB0.7 was localised by somatic cell Chromosomal Localisation: Chromosome 17pl2-pl3 (3). hybrids and pulsed field gel electrophoresis to 19ql3.3, approximately 150 kb distal to D19S63. The locus is now Mendelian Inheritance: Co-dominant segregation shown in three designated D19S95. informative two generation families. Mendelian Inheritance: Co-dominant segregation has been Probe Availability: Requests should be sent to Professor observed in 38 families. R.I.Handin, Brigham and Women's Hospital, Hematology Probe Availability: Available for collaborative studies. Contact Dr D.J.Shaw. Comments: The CTG repeat associated with myotonic dystrophy (DM) is located 9.5 kb from pBB0.7. This repeat is known to expand in individuals with DM. Expansion of the repeat can be detected when pBB0.7 is hybridised to genomic DNA digested with EcoRl. The expansion is always associated with the 10 kb EcoRl allele (2, 3). Acknowledgements: We wish to thank the patients and their families; the UK Muscular Dystrophy Group, the Muscular Dystrophy Association and the Piton Foundation (USA), and the Wellcome Trust for financial support; colleagues in the Myotonic Dystrophy Working Group; and L.A.Sandkuijl for advice on statistical analysis.
Division, 75 Francis Street, Boston, MA 02115, USA. Other Comments: The polymorphic Taql site was mapped and sequenced. It is located 75 bp 3' of the 2356 bp Gplba exon, which contains the complete coding sequence of the gene. The finding of the polymorphisms may be of use in the study of patients with various forms of the Bernard-Soulier Syndrome, a rare inherited bleeding disorder which involves the GPIB/IX complex on platelets. Acknowledgement: This study was supported by NIH grant ROI HL 34787.
References: 1) Brook,J.D. et al. (1992) Genomics 13, 243-250. 2) Harley,H.G. et al. (1992) Nature 355, 545-546. 3) Brook.J.D. et al. (1992) Cell 68, 799-808.
References: 1) Lopez.J.A., Chung,D.W., Fujikawa,K., Hagen.F.S. Papayannopoulou,T. and Roth,G.J. (1987) Proc. Natl. Acad. Sci. USA 84, 5615-5619. 2) Cruz.M.A. Petersen.E.J. and Handin.R.I. (1992) J. Biol. Chem. 276, 1303-1309. 3) Wenger,R.H., Wicki.A.N., Kieffer.N., Adolph,S., Hameister,H. and Clemetson.K.J. (1989) Gene 85, 517-524.
* To whom correspondence should be addressed
•To whom correspondence should be addressed at: University Hospital, Utrecht, Department of Hematology, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands
Downloaded from http://hmg.oxfordjournals.org/ at University of Chicago on July 3, 2015
Source/Description: DNA probe pBB0.7 is a 700 bp BamHl fragment derived from a lambda library created from 2F5 — a hybrid cell line containing the myotonic dystrophy region of chromosome 19 as its only human material (1).
Source/Description: Southern blotting was used to detect two polymorphic sites in the human glycoprotein Iba (GPIBa) gene. Full length GPIBa cDNA, subcloned in the EcoRl site of pUC 19 was used as a probe (1, 2).
Insertion/deletion polymorphism at D19S95 associated with the myotonic dystrophy CTG repeat
Taq\ and Bsu36\ polymorphisms in the human glycoprotein Iba gene (GPIBa:)
S.R.Crow*, H.G.Harley, J.D.Brook1, S.A.Rundle and D.J.Shaw Institute of Medical Genetics, University of Wales College of Medicine, Heath Park, Cardiff, CF4 6EJ, UK and 1 Centre for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
E.J.Petersen* and R.I.Handin Brigham and Women's Hospital, Department of Hematology and Harvard Medical School, Department of Medicine, Boston, MA, USA
Polymorphism: pBB0.7 identifies a 1.0 kb insertion/deletion RFLP with the enzymes EcoRV (10.5 kb, 9.5 kb), Eco9A (10 kb, 9 kb), HincJl (14 kb, 13 kb), PvuU (5.0 kb, 4.0 kb) and BgK (5.0 kb, 4.0 kb). Frequency: For the EcoRV RFLP, allele frequencies studied in 90 unrelated individuals are as follows: Al (10.5) 0.56 A2 (9.5) 0.44 Heterozygosity = 0.49.
Polymorphisms: Taql identifies a polymorphic site, detected as the presence or absence of a 4.0 kb fragment, with constant bands at 0.5, 0.7 and 1.1 kb. Bsu36I identifies a polymorphic site, detected as the presence or absence of a 6.9 kb fragment, with constant bands at 6.0 kb. Frequency: Taql studied in ten unrelated Caucasian individuals and Bsu36I in 15. Taql
+ 0.15 - 0.85 Heterozygosity = 0.26
Bsu36I
+ 0.20 - 0.80 Heterozygosity = 0.32
Not Polymorphic For: AccI, Apal, BamHI, Bell, BgU, Bgin, Not Polymorphic For: Aval, BamHl, Banl, BgH, HinfU, HindSR, Dral, EcoRl, EcoRV, HaeDI, HindHI, Hinfl, Hpafl, Kpnl, Ncol, Pstl, PvuE, Sad, Sau3AI, Xbal, XmnI, as studied in 10 unrelated HaeUl, Mspl, Pstl, Rsal, Sail, Smal, Sphl, Taql, Xbal. Caucasians. Chromosomal Localisation: pBB0.7 was localised by somatic cell Chromosomal Localisation: Chromosome 17pl2-pl3 (3). hybrids and pulsed field gel electrophoresis to 19ql3.3, approximately 150 kb distal to D19S63. The locus is now Mendelian Inheritance: Co-dominant segregation shown in three designated D19S95. informative two generation families. Mendelian Inheritance: Co-dominant segregation has been Probe Availability: Requests should be sent to Professor observed in 38 families. R.I.Handin, Brigham and Women's Hospital, Hematology Probe Availability: Available for collaborative studies. Contact Dr D.J.Shaw. Comments: The CTG repeat associated with myotonic dystrophy (DM) is located 9.5 kb from pBB0.7. This repeat is known to expand in individuals with DM. Expansion of the repeat can be detected when pBB0.7 is hybridised to genomic DNA digested with EcoRl. The expansion is always associated with the 10 kb EcoRl allele (2, 3). Acknowledgements: We wish to thank the patients and their families; the UK Muscular Dystrophy Group, the Muscular Dystrophy Association and the Piton Foundation (USA), and the Wellcome Trust for financial support; colleagues in the Myotonic Dystrophy Working Group; and L.A.Sandkuijl for advice on statistical analysis.
Division, 75 Francis Street, Boston, MA 02115, USA. Other Comments: The polymorphic Taql site was mapped and sequenced. It is located 75 bp 3' of the 2356 bp Gplba exon, which contains the complete coding sequence of the gene. The finding of the polymorphisms may be of use in the study of patients with various forms of the Bernard-Soulier Syndrome, a rare inherited bleeding disorder which involves the GPIB/IX complex on platelets. Acknowledgement: This study was supported by NIH grant ROI HL 34787.
References: 1) Brook,J.D. et al. (1992) Genomics 13, 243-250. 2) Harley,H.G. et al. (1992) Nature 355, 545-546. 3) Brook.J.D. et al. (1992) Cell 68, 799-808.
References: 1) Lopez.J.A., Chung,D.W., Fujikawa,K., Hagen.F.S. Papayannopoulou,T. and Roth,G.J. (1987) Proc. Natl. Acad. Sci. USA 84, 5615-5619. 2) Cruz.M.A. Petersen.E.J. and Handin.R.I. (1992) J. Biol. Chem. 276, 1303-1309. 3) Wenger,R.H., Wicki.A.N., Kieffer.N., Adolph,S., Hameister,H. and Clemetson.K.J. (1989) Gene 85, 517-524.
* To whom correspondence should be addressed
•To whom correspondence should be addressed at: University Hospital, Utrecht, Department of Hematology, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands
Downloaded from http://hmg.oxfordjournals.org/ at University of Chicago on July 3, 2015
Source/Description: DNA probe pBB0.7 is a 700 bp BamHl fragment derived from a lambda library created from 2F5 — a hybrid cell line containing the myotonic dystrophy region of chromosome 19 as its only human material (1).
Source/Description: Southern blotting was used to detect two polymorphic sites in the human glycoprotein Iba (GPIBa) gene. Full length GPIBa cDNA, subcloned in the EcoRl site of pUC 19 was used as a probe (1, 2).
