Population study of a sequence polymorphism in intron 2 of the human ?-globin gene

Braun, Andreas; Ambach, Helmut; Bichlmaier, Regina; Kammerer, Stefan; Aa, Roscher

doi:10.1007/bf00225207

Cited by 8 publications

(5 citation statements)

References 1 publication

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“…The general applicability of the "selected conditions" was examined by performing SSCP analyses of a 233 base pair fragment specific for intron 2 of the human -globin gene (amplification as described by Braun et al, (11)) and of segments of 202, 213, 239 and 227 base pairs of exons 1, 5, 6 and 9 of the gene responsible for X-chromosomal adrenoleukodystrophy (amplification conditions and the sequences of these novel mutants will be published elsewhere in detail).…”

Section: Dna Segments Of the Human -Globin Gene And The Gene Responsimentioning

confidence: 99%

Conditions for Single Strand Conformation Polymorphism (SSCP) Analysis with Broad Applicability: A Study on the Effects of Acrylamide, Buffer and Glycerol Concentrations in SSCP Analysis of Exons of the p53 Gene

Teschauer¹,

Mussack²,

Braun³

et al. 1996

CCLM

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We examined the influence of electrophoretic conditions on the detectability of small sequence alterations in DNA fragments by single strand conformation polymorphism (SSCP) analysis. Three acrylamide concentrations, 7.5, 14 and 20%, were selected, and all three gel types were prepared with four different Tris/borate/EDTA buffer concentrations. In addition, all these twelve gels were prepared both with and without 10% glycerol. All electrophoretic runs were performed at ambient temperature of 20-24 °C. The resulting 24 different electrophoretic conditions were used for the SSCP analysis of DNA fragments of exons 5, 7 and 8 of the human p53 gene; the results were evaluated primarily in relation to detectability of mutants. Six out of the 24 conditions permitted the detection of all mutants. For practical reasons, 14% acrylamide, 44.5 mmol/1 Tris, 44.5 mmol/1 boric acid, 1 mmol/1 EDTA, pH 8.0, with and without addition of glycerol was chosen as the most suitable. These "selected conditions" were applied in the SSCP analysis of an arbitrarily chosen set of mutant DNA fragments with single base exchanges, and all but one of seven mutants were detected in the gel system containing glycerol.Our results indicate that the set of "selected conditions" is of broad applicability, permitting the detection of even small sequence differences like single base exchanges with high reliability. It should prove especially useful in screening for unknown mutations.

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Section: Dna Segments Of the Human -Globin Gene And The Gene Responsimentioning

confidence: 99%

Conditions for Single Strand Conformation Polymorphism (SSCP) Analysis with Broad Applicability: A Study on the Effects of Acrylamide, Buffer and Glycerol Concentrations in SSCP Analysis of Exons of the p53 Gene

Teschauer¹,

Mussack²,

Braun³

et al. 1996

CCLM

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“…The adrenoleukodystrophy protein (ALDP) gene has been cloned, and it was thought to be a disease-causing candidate for X-ALD (10). Various point mutations, deletions, and insertions in the gene have been identified in these patients (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21). Retroviral transfer of normal ALDP cDNA into patient's fibroblasts enhanced VLCFA oxidation activity to the level found in control fibroblasts (22).…”

mentioning

confidence: 99%

Molecular Cloning of cDNA Encoding Rat Very Long-chain Acyl-CoA Synthetase

Uchiyama

Aoyama

Kamijo

et al. 1996

Journal of Biological Chemistry

147

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The cDNA encoding rat very long-chain acyl-CoA synthetase (VLACS) was cloned, using degenerative primers synthesized according to the partial amino acid sequences of the peptide fragments of the purified rat liver enzyme. The longest cDNA insert was 2972 base pairs with a 1860-base pair open reading frame encoding 620 amino acids. The calculated molecular mass of 70,692 daltons was consistent with size of the purified enzyme. In Northern blot analysis, a single band was detected at the position of about 3 kilobases, corresponding to the size of the cloned cDNA. cDNA-directed expression in Escherichia coli resulted in accumulation of expressed protein, as an inclusion body. An antibody was raised using this expressed protein to characterize the cDNA and the enzyme. The subcellular localization of VLACS in peroxisomes and microsomes was demonstrated in Western blot analysis. The specific activity and the substrate specificity of the cDNA expressed enzyme in COS-1 cells were consistent with those of the purified rat enzyme. The predicted amino acid sequence of VLACS had a high sequence similarity to fatty acid transport protein (Schaffer, J. E., and Lodish, H. F. (1994) Cell 79, 427-436), and was considered to have domains for adenylation and thioester formation. The entire structure of VLACS was dissimilar to that of longchain acyl-CoA synthetase

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“…The frequencies of the less common alleles of these three SNPs were found to be 0.436 for CD2 T, 0.446 for IVS2 ϩ 666 C, and 0.010 for IVS2 ϩ 672 C in normal Chinese individuals. It is interesting to note the large difference of the allele frequencies of IVS2 ϩ 666 C in Chinese (0.446), Caucasians (0.14), and Africans (0.13) (Braun et al, 1995).…”

Section: Normal Sscp Patternsmentioning

confidence: 99%

Rapid Detection of Common Southeast Asian ß-Thalassemia Mutations by Nonisotopic Multiplex PCR-SSCP Analysis

Yip

Fung²,

Lo³

2004

Genetic Testing

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This report describes the detection of seven beta-thalassemia mutations common in Southeast Asia by amplifying three short PCR fragments in two separate tubes, followed by single-strand conformation polymorphism (SSCP) analysis in single lanes. These mutations are -28 A --> G, codon 17 A --> T, IVS1 + 5 G --> C, codon 41/42 -CTTT, codon 43 G --> T, codon 71/72 + A, and IVS2 + 654 C --> T, and account for 70% to over 95% of the cases in this region. This rapid nonisotopic method was also found capable of detecting other mutations within the amplified fragments. It is simple, rapid, and cheap, and thus suitable for carrier screening and prenatal diagnosis in Southeast Asia.

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Population study of a sequence polymorphism in intron 2 of the human ?-globin gene

Cited by 8 publications

References 1 publication

Conditions for Single Strand Conformation Polymorphism (SSCP) Analysis with Broad Applicability: A Study on the Effects of Acrylamide, Buffer and Glycerol Concentrations in SSCP Analysis of Exons of the p53 Gene

Conditions for Single Strand Conformation Polymorphism (SSCP) Analysis with Broad Applicability: A Study on the Effects of Acrylamide, Buffer and Glycerol Concentrations in SSCP Analysis of Exons of the p53 Gene

Molecular Cloning of cDNA Encoding Rat Very Long-chain Acyl-CoA Synthetase

Rapid Detection of Common Southeast Asian ß-Thalassemia Mutations by Nonisotopic Multiplex PCR-SSCP Analysis

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