Nucleic Acids Research, Vol. 19, No. 5 1163
Dinucleotide repeat polymorphism in the human glucagon gene (GCG)
A new polymorphic probe on chromosome 3p: XLIB41 -51 (D3S632)
S.Wu, K.Xiang and G.l.Bell Howard Hughes Medical Institute, University of Chicago, 5841 S. Maryland Ave, Box 391, Chicago, IL 60637, USA
F.Latif, G.M.Glenn, K.Tory, H.Brauch, J.Delisio1, K.Hampsch1, M.L.Orcutt, B.Zbar and M.l.Lerman* Laboratory of Immunobiology, National Cancer Institute- Frederick Cancer Research and Development Center, Frederick, MD 21701 and 1Progam Resources Inc., Frederick Cancer Research and Development Center, Frederick, MD 21701, USA
Source/Description: Two primers (GCG-1, AAAGGACATAGAT'lTGTCTG, and GCG-2, GCAACTCTGT'lTCTCCTATC) were used to amplify a 125 bp GA repeat-rich region in intron 2 of the human glucagon gene (GCG) (1, 2). Frequency: Eight alleles (numbered in order of decreasing size) were observed in 34 unrelated Caucasians. The heterozygosity was 82%. Allele Frequency 0.32 A1 A2 0.15 A3 0.24 A4 0.13 A5 0.03 A6 0.06 A7 0.06 A8 0.01 Chromosomal Localization: GCG has been localized to chromosome 2q36 -q37 (3). Mendelian Inheritance: Co-dominant inheritance was observed in a five-generation family in which 50 individuals were typed. Other Comments: The PCR was performed using 32P-labeled GCG-1 and unlabeled GCG-2 for 30 cycles: denaturation at 94°C for 1 min; annealing at 55°C for 2 min; and extension at 72°C for 3 min. The PCR products were analyzed on a 5 % denaturing polyacrylamide gel (Fig. 1). References: 1) Bell,G.I. et al. (1983) Nature 304, 368. 2) White,J.W. and Saunders,G.F. (1986) Nucl. Acids Res. 14, 4719. 3) Schroeder,W.T. et al. (1984) Cytogenet. Cell Genet. 38, 76.
Source/Description: XLIB41 -51 is a 4.3 kb EcoRI fragment isolated from a Charon 21A human chromosome 3 library (1). Polymorphism: MspI digestion of genomic DNA and hybridization with the probe detects a two allele polymorphism: 8.0 kb (Al) and 6.6 kb with 1.4 kb (A2). Frequency: Estimated from 120 unrelated Caucasians. Al : 0.61 A2 : 0.39 Frequency of Heterozygosity: 0.48 Not Polymorphic For: BamHI, DraI, EcoRI, MspI, RsaI, BglII,
HinfI, TaqI and Pvul. Chromosomal Localization: Using a somatic cell hybrid panel (2) which was based on linkage groups anchored to physically localized markers, the probe was assigned to the short arm of chromosome 3 between 3p22 -3p26. Mendelian Inheritance: Mendelian inheritance has been demonstrated in a three generation CEPH family. Probe Availability: Available for collaboration. References: 1) American Type Culture Collection No. 57748, National Lab ID Code LA03NS02. 2) Lerman,M., Latif,F., Glenn,G. et al.: Isolation and regional localization of a large collection (2,000) of single copy DNA fragments on human chromosome 3 for mapping and cloning tumor suppressor genes. Human Genetics (In press, 1990).
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Figure 1. PCR amplification of GCG dinucleotide repeat DNA polymorphism. The genotypes of the seven individuals shown are (left to right): 35, 23, 25, 35, 15, 23 and 15.
*To whom correspondence should be addressed at Laboratory of Immunology, Frederick Cancer Research and Development Center, NCINIH, Ft. Detrick, Bldg 560, Rm 12-71, Frederick, MD 21701, USA
Dinucleotide repeat polymorphism in the human glucagon gene (GCG)
A new polymorphic probe on chromosome 3p: XLIB41 -51 (D3S632)
S.Wu, K.Xiang and G.l.Bell Howard Hughes Medical Institute, University of Chicago, 5841 S. Maryland Ave, Box 391, Chicago, IL 60637, USA
F.Latif, G.M.Glenn, K.Tory, H.Brauch, J.Delisio1, K.Hampsch1, M.L.Orcutt, B.Zbar and M.l.Lerman* Laboratory of Immunobiology, National Cancer Institute- Frederick Cancer Research and Development Center, Frederick, MD 21701 and 1Progam Resources Inc., Frederick Cancer Research and Development Center, Frederick, MD 21701, USA
Source/Description: Two primers (GCG-1, AAAGGACATAGAT'lTGTCTG, and GCG-2, GCAACTCTGT'lTCTCCTATC) were used to amplify a 125 bp GA repeat-rich region in intron 2 of the human glucagon gene (GCG) (1, 2). Frequency: Eight alleles (numbered in order of decreasing size) were observed in 34 unrelated Caucasians. The heterozygosity was 82%. Allele Frequency 0.32 A1 A2 0.15 A3 0.24 A4 0.13 A5 0.03 A6 0.06 A7 0.06 A8 0.01 Chromosomal Localization: GCG has been localized to chromosome 2q36 -q37 (3). Mendelian Inheritance: Co-dominant inheritance was observed in a five-generation family in which 50 individuals were typed. Other Comments: The PCR was performed using 32P-labeled GCG-1 and unlabeled GCG-2 for 30 cycles: denaturation at 94°C for 1 min; annealing at 55°C for 2 min; and extension at 72°C for 3 min. The PCR products were analyzed on a 5 % denaturing polyacrylamide gel (Fig. 1). References: 1) Bell,G.I. et al. (1983) Nature 304, 368. 2) White,J.W. and Saunders,G.F. (1986) Nucl. Acids Res. 14, 4719. 3) Schroeder,W.T. et al. (1984) Cytogenet. Cell Genet. 38, 76.
Source/Description: XLIB41 -51 is a 4.3 kb EcoRI fragment isolated from a Charon 21A human chromosome 3 library (1). Polymorphism: MspI digestion of genomic DNA and hybridization with the probe detects a two allele polymorphism: 8.0 kb (Al) and 6.6 kb with 1.4 kb (A2). Frequency: Estimated from 120 unrelated Caucasians. Al : 0.61 A2 : 0.39 Frequency of Heterozygosity: 0.48 Not Polymorphic For: BamHI, DraI, EcoRI, MspI, RsaI, BglII,
HinfI, TaqI and Pvul. Chromosomal Localization: Using a somatic cell hybrid panel (2) which was based on linkage groups anchored to physically localized markers, the probe was assigned to the short arm of chromosome 3 between 3p22 -3p26. Mendelian Inheritance: Mendelian inheritance has been demonstrated in a three generation CEPH family. Probe Availability: Available for collaboration. References: 1) American Type Culture Collection No. 57748, National Lab ID Code LA03NS02. 2) Lerman,M., Latif,F., Glenn,G. et al.: Isolation and regional localization of a large collection (2,000) of single copy DNA fragments on human chromosome 3 for mapping and cloning tumor suppressor genes. Human Genetics (In press, 1990).
r4
Figure 1. PCR amplification of GCG dinucleotide repeat DNA polymorphism. The genotypes of the seven individuals shown are (left to right): 35, 23, 25, 35, 15, 23 and 15.
*To whom correspondence should be addressed at Laboratory of Immunology, Frederick Cancer Research and Development Center, NCINIH, Ft. Detrick, Bldg 560, Rm 12-71, Frederick, MD 21701, USA
