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PubMed · 9360636

Search for DNA sequence variations using a MutS-based technology.

C Bellanné-Chantelot·S Beaufils·V Hourdel·S Lesage·V Morel·N Dessinais·I Le Gall·D Cohen·J Dausset
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Abstract

The search for DNA sequence variations (DSV) is emphasized with genetic studies of a large number of multifactorial diseases. Saturation of regions of interest with diallelic polymorphisms will be an essential step to pinpoint, through association studies, predisposing genes. We have developed a solid-phase method based on the ability of mismatch binding protein MutS to recognize single nucleotide mismatches. This approach was applied to the study of 83 sequence-tagged sites (STSs) extracted from an eight centimorgans (cM) chromosome 21 region. One-third of tested STSs were found to be polymorphic leading to a frequency of one DSV every 822 base pairs (bp). Sequencing of analyzed STSs showed the high reliability of the MutS-based technology for mismatches up to 2 bp in DNA fragments ranging in size from 200 bp to 1 kilobase (kb). The entire assay which is performed in a solid-phase format without the need of electrophoresis or sequencing, will provide an efficient tool for new polymorphism detection.

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Adenosine TriphosphatasesBacterial ProteinsBase SequenceChromosome MappingChromosomes, Human, Pair 21DNADNA-Binding ProteinsEscherichia coli ProteinsGenetic VariationHumansMolecular Probe TechniquesMutS DNA Mismatch-Binding ProteinNucleic Acid HeteroduplexesPolymorphism, GeneticSequence Analysis, DNASequence Tagged SitesJournal ArticleResearch Support, Non-U.S. Gov't

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C Bellanné-Chantelot, S Beaufils, V Hourdel, S Lesage, V Morel, N Dessinais, I Le Gall, D Cohen, J Dausset. 1997. Search for DNA sequence variations using a MutS-based technology.. https://doi.org/10.1016/s1383-5726(97)00007-1

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