Nucleic Acids Research, Vol. 19, No. 18 5095
Length variation within intron 2 of the human IL-1 receptor antagonist protein gene (ILl RN) A.Steinkasserer, K.Koelble and R.B.Sim MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK
Source/Description: The PCR primers 5'- GTTGCTGGATACTTGCAAGGACCA-3' and 5'- CCCTCCATGGATTCCCAAGAACAG-3' were used to amplify the 2nd intron of IL1RN (1) in a Cetus Thermal cycler (denaturing temp. 93°C-30 sec, annealing temp. 68°C-30 sec, extension temp. 72°C-5 minutes). The products were analyzed in a 1% (w/v) agarose gel. Polymorphism: A triallelic polymorphism was detected with the following size variations: allele Al (1.55 kb), allele A2 (1.45 kb) and allele A3 (1.3 kb). The phenotypes A1/A2, A1/A3, A2/A3, A2/A2 and A3/A3 were observed in a sample of 107 individuals (as illustrated in Fig. 1). Frequency: In a random laboratory population the following allele frequencies were observed: Al 0.04 A2 0.65 A3 0.30 (Chromosomes n = 78). Mendelian Inheritance: For the A2 and A3 alleles co-dominant segregation was observed in a three generation and in four twogeneration families. Due to the rarity of the Al allele segregation could not be found. Other Comments: Using an extension time of 3 min at 72°C intermediate-sized products of 1.4 and 1.5 kb were observed in A2A3 and A1A3 heterozygotes, respectively. These PCR artefacts are probably due to in vitro recombination or template strand switching (2). These artefacts were not detected when the extension time at 72°C was increased to 5 min. Acknowledgements: This work was a contribution from the Oxford Centre for Molecular Science (OCMS) which is supported by the SERC and the MRC. References: 1) Eisenberg et al. (1991) PNAS 88, 5232-5236. 2) Erlich et al. (1991) Science 252, 1643-1651.
A highly polymorphic probe on 11 p15.5: L22.5.2 (D11 S774) A.Puech, I.Henry, C.Jeanpierre and C.Junien* INSERM U73, Chateau de Longchamp, Carrefour de Longchamp, Bois de Boulogne, 75016 Paris, France
Source/Description: L22.5.2 (Dl 1S774) is a 19 kb MboI fragment isolated from an EMBL3 human chromosome lpl5.5 specific library. Polymorphism: MspI digests of genomic DNA identify a multiple allele polymorphism with at least six bands of 3.8 kb (allele 1), 3.5 kb (allele 2), 3.25 kb (allele 3), 3.1 kb (allele 4), 2.95 kb (allele 5) and 2.55 kb (allele 6). NcoI, PvuII, TaqI also identify the same polymorphism (six individuals) with bands ranging from 6.4 to 7.5 kb, 4.3 to 5.2 kb and 6.4 to 7.8 kb respectively. Allele Frequency: Frequencies of the alleles are based on the MspI digests of DNAs from 70 unrelated European Caucasians Al: 0.124 A2: 0.194 A3: 0.202 A4: 0.178 A5: 0.225 A6: 0.077 Not Polymorphic For: HindIII, EcoRV, BamHI, BglII, EcoRI on six individuals. Chromosomal Localization: Using a somatic cell hybrid panel of chromosome 11 (1), the probe was localized in I lplS.5, distal to Dl S26. Mendelian Inheritance: Codominant segregation was observed in a three generation family. Hybridization Conditions: The probe was pre-associated with excess total human DNA prior to hybridization (2). Probe Availability: Available for collaboration. Acknowledgements: AP is supported by a grant from the Ministere de la Recherche et de la Technologie. This work was supported by grants from the Ligue Nationale contre le Cancer, Association pour la Recherche contre le Cancer, INSERM and Faculte de Medecine Paris-Ouest. References: 1) Glaser,T. et al. (1989) Somatic Cell. Mol. Genet. 15, 477-501. 2) Nakamura,Y. et al. (1988) Am. J. Hum. Genet. 43, 854-859.
*
To whom
correspondence should be addressed
Length variation within intron 2 of the human IL-1 receptor antagonist protein gene (ILl RN) A.Steinkasserer, K.Koelble and R.B.Sim MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK
Source/Description: The PCR primers 5'- GTTGCTGGATACTTGCAAGGACCA-3' and 5'- CCCTCCATGGATTCCCAAGAACAG-3' were used to amplify the 2nd intron of IL1RN (1) in a Cetus Thermal cycler (denaturing temp. 93°C-30 sec, annealing temp. 68°C-30 sec, extension temp. 72°C-5 minutes). The products were analyzed in a 1% (w/v) agarose gel. Polymorphism: A triallelic polymorphism was detected with the following size variations: allele Al (1.55 kb), allele A2 (1.45 kb) and allele A3 (1.3 kb). The phenotypes A1/A2, A1/A3, A2/A3, A2/A2 and A3/A3 were observed in a sample of 107 individuals (as illustrated in Fig. 1). Frequency: In a random laboratory population the following allele frequencies were observed: Al 0.04 A2 0.65 A3 0.30 (Chromosomes n = 78). Mendelian Inheritance: For the A2 and A3 alleles co-dominant segregation was observed in a three generation and in four twogeneration families. Due to the rarity of the Al allele segregation could not be found. Other Comments: Using an extension time of 3 min at 72°C intermediate-sized products of 1.4 and 1.5 kb were observed in A2A3 and A1A3 heterozygotes, respectively. These PCR artefacts are probably due to in vitro recombination or template strand switching (2). These artefacts were not detected when the extension time at 72°C was increased to 5 min. Acknowledgements: This work was a contribution from the Oxford Centre for Molecular Science (OCMS) which is supported by the SERC and the MRC. References: 1) Eisenberg et al. (1991) PNAS 88, 5232-5236. 2) Erlich et al. (1991) Science 252, 1643-1651.
A highly polymorphic probe on 11 p15.5: L22.5.2 (D11 S774) A.Puech, I.Henry, C.Jeanpierre and C.Junien* INSERM U73, Chateau de Longchamp, Carrefour de Longchamp, Bois de Boulogne, 75016 Paris, France
Source/Description: L22.5.2 (Dl 1S774) is a 19 kb MboI fragment isolated from an EMBL3 human chromosome lpl5.5 specific library. Polymorphism: MspI digests of genomic DNA identify a multiple allele polymorphism with at least six bands of 3.8 kb (allele 1), 3.5 kb (allele 2), 3.25 kb (allele 3), 3.1 kb (allele 4), 2.95 kb (allele 5) and 2.55 kb (allele 6). NcoI, PvuII, TaqI also identify the same polymorphism (six individuals) with bands ranging from 6.4 to 7.5 kb, 4.3 to 5.2 kb and 6.4 to 7.8 kb respectively. Allele Frequency: Frequencies of the alleles are based on the MspI digests of DNAs from 70 unrelated European Caucasians Al: 0.124 A2: 0.194 A3: 0.202 A4: 0.178 A5: 0.225 A6: 0.077 Not Polymorphic For: HindIII, EcoRV, BamHI, BglII, EcoRI on six individuals. Chromosomal Localization: Using a somatic cell hybrid panel of chromosome 11 (1), the probe was localized in I lplS.5, distal to Dl S26. Mendelian Inheritance: Codominant segregation was observed in a three generation family. Hybridization Conditions: The probe was pre-associated with excess total human DNA prior to hybridization (2). Probe Availability: Available for collaboration. Acknowledgements: AP is supported by a grant from the Ministere de la Recherche et de la Technologie. This work was supported by grants from the Ligue Nationale contre le Cancer, Association pour la Recherche contre le Cancer, INSERM and Faculte de Medecine Paris-Ouest. References: 1) Glaser,T. et al. (1989) Somatic Cell. Mol. Genet. 15, 477-501. 2) Nakamura,Y. et al. (1988) Am. J. Hum. Genet. 43, 854-859.
*
To whom
correspondence should be addressed
