Search for DNA sequence variations using a MutS-based technology.
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.
Bellanné-Chantelot
C
C
Fondation Jean Dausset-CEPH, Paris, France.
Beaufils
S
S
Hourdel
V
V
Lesage
S
S
Morel
V
V
Dessinais
N
N
Le Gall
I
I
Cohen
D
D
Dausset
J
J
D000251:Adenosine Triphosphatases; D001426:Bacterial Proteins; D001483:Base Sequence; D002874:Chromosome Mapping; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics; D004247:DNA / Q000235:genetics*; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D014644:Genetic Variation / Q000235:genetics*; D006801:Humans; D015336:Molecular Probe Techniques; D051719:MutS DNA Mismatch-Binding Protein; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA; D016324:Sequence Tagged Sites
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D009692:Nucleic Acid Heteroduplexes; D004247:DNA; D000251:Adenosine Triphosphatases; D051719:MutS DNA Mismatch-Binding Protein; C497175:MutS protein, E coli