Rapid Genotyping of the Osteoporosis-Associated Polymorphic Transcription Factor Sp1 Binding Site in the COL1A1 Gene by Pyrosequencing
| Autor: | Katarina Ellnebo-Svedlund, Lasse Larsson, Per Magnusson, Jon Jonasson |
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| Rok vydání: | 2004 |
| Předmět: |
Genotype
Sp1 Transcription Factor Bioengineering Single-nucleotide polymorphism Biology Polymerase Chain Reaction Polymorphism Single Nucleotide Applied Microbiology and Biotechnology Biochemistry Collagen Type I law.invention Bone Density law Humans Molecular Biology Gene Genotyping Alleles Polymerase chain reaction Genetics Binding Sites Polymorphism Genetic Sequence Analysis DNA Amplicon Molecular biology Genetic Techniques Genetic marker Osteoporosis Pyrosequencing Biotechnology |
| Zdroj: | Molecular Biotechnology. 26:87-90 |
| ISSN: | 1073-6085 |
| DOI: | 10.1385/mb:26:1:87 |
| Popis: | We describe a novel polymerase chain reaction (PCR) and deoxyribonucleic acid (DNA) sequencingbased assay for rapid genotyping of the polymorphic Sp1 binding site in the COL1A1 gene (1). A single nucleotide G--T substitution polymorphism at this GC-rich site has recently been reported to be a predictive genetic marker for low bone mineral density (BMD). To simplify screening for this marker, we optimized PCR conditions and subjected the amplicons to pyrosequencing, which is a convenient high-throughput sequence analysis technique, readily amenable to automation. The analysis of 200 deidentified convenience DNA samples extracted from blood revealed genotype frequences in Hardy-Weinberg equilibrium (SS 68.0%, Ss 28.5%, and ss 3.5%) in agreement with other studies of European populations. This study demonstrates for the first time that pyrosequencing can be used for rapid identification of the osteoporosis-associated single nucleotide polymorphism (SNP) in the COL1A1 gene. |
| Databáze: | OpenAIRE |
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