Detection of multiallele polymorphisms within gene sequences by GC-clamped denaturing gradient gel electrophoresis.
The ability to efficiently detect DNA polymorphisms is essential for the completion of a high-resolution polymorphic linkage map of the human genome. Currently the most informative polymorphisms are the multiallelic dinucleotide repeat polymorphisms. However, many gene sequences lack an associated dinucleotide repeat sequence. We used GC-clamped denaturing gradient gel electrophoresis to screen for DNA polymorphisms in the following six gene sequences: MCC, p53, prealbumin (transthyretin), rhodopsin, S-antigen, and TGF-alpha. A single-base sequence polymorphism was identified in each of these gene sequences. Some of these polymorphisms were multiallelic and highly informative. Our results demonstrate the value of denaturing gradient gel electrophoresis for both identifying and analyzing human DNA polymorphisms. The ability to detect highly informative polymorphisms within gene sequences will greatly contribute to a gene-based polymorphic linkage map.
Sheffield
V C
VC
Department of Pediatrics University of Iowa, Iowa City.
Beck
J S
JS
Nichols
B
B
Cousineau
A
A
Lidral
A C
AC
Stone
E M
EM
D000483:Alleles; D000941:Antigens / Q000235:genetics; D019393:Arrestin; D001483:Base Sequence; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D005136:Eye Proteins / Q000235:genetics; D016147:Genes, Tumor Suppressor / Q000235:genetics; D016158:Genes, p53 / Q000235:genetics; D008040:Genetic Linkage / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic / Q000235:genetics*; D011228:Prealbumin / Q000235:genetics; D011506:Proteins / Q000235:genetics; D012243:Rhodopsin / Q000235:genetics; D016211:Transforming Growth Factor alpha / Q000235:genetics; D025521:Tumor Suppressor Proteins
D000941:Antigens; D019393:Arrestin; D005136:Eye Proteins; D011228:Prealbumin; D011506:Proteins; D016211:Transforming Growth Factor alpha; D025521:Tumor Suppressor Proteins; C067857:MCC protein, human; D004247:DNA; D012243:Rhodopsin