Identification of novel rhodopsin mutations associated with retinitis pigmentosa by GC-clamped denaturing gradient gel electrophoresis

Val C Sheffield; Gerald A Fishman; John S Beck; Alan E Kimura; Edwin M Stone

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Identification of novel rhodopsin mutations associated with retinitis pigmentosa by GC-clamped denaturing gradient gel electrophoresis

Journal article

Peer reviewed

Identification of novel rhodopsin mutations associated with retinitis pigmentosa by GC-clamped denaturing gradient gel electrophoresis

Val C Sheffield, Gerald A Fishman, John S Beck, Alan E Kimura and Edwin M Stone

American journal of human genetics, Vol.49(4), pp.699-706

10/1991

PMCID: PMC1683182

PMID: 1897520

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Abstract

Retinitis pigmentosa (RP) is a group of disorders characterized by progressive degeneration of the outer retina, resulting in night blindness, visual field loss, an abnormal electroretinogram, and characteristic retinal pigmentary changes. An important step in the understanding of RP has been the recognition that some cases of autosomal dominant RP (ADRP) are caused by mutations in the rhodopsin gene. Multiple different point mutations within the coding sequence of the rhodopsin gene have been associated with ADRP. We have developed a GC-clamped denaturing-gradient-gel electrophoresis (DGGE) assay for the coding region of the rhodopsin gene and have used this assay to screen ADRP patients for mutations. The assay consists of amplifying with PCR the five exons of the rhodopsin gene and then analyzing each PCR product by DGGE. We have used this assay to detect three previously unreported rhodopsin base substitutions associated with ADRP. The use of this assay to identify ADRP patients who have various rhodopsin mutations has allowed us to begin studies seeking to correlate molecular genotype with clinical phenotype. Furthermore, GC-clamped DGGE has allowed us to identify families with ADRP not caused by a rhodopsin mutation. Such families will be important in the search for other genes involved in ADRP.

Polymerase Chain Reaction

Amino Acid Sequence

Electrophoresis, Polyacrylamide Gel

Humans

Molecular Sequence Data

Retinitis Pigmentosa - genetics

Exons - genetics

Male

Mutation - genetics

Genetic Testing - methods

Polymorphism, Genetic

Rhodopsin - genetics

Oligodeoxyribonucleotides - genetics

Pedigree

Base Sequence

Female

Details

Title: Subtitle: Identification of novel rhodopsin mutations associated with retinitis pigmentosa by GC-clamped denaturing gradient gel electrophoresis
Creators: Val C Sheffield - Department of Pediatrics, University of Iowa, Iowa City
Gerald A Fishman
John S Beck
Alan E Kimura
Edwin M Stone
Resource Type: Journal article
Publication Details: American journal of human genetics, Vol.49(4), pp.699-706
PMID: 1897520
PMCID: PMC1683182
NLM abbreviation: Am J Hum Genet
ISSN: 0002-9297
eISSN: 1537-6605
Publisher: United States
Grant note: EYO8426 / NEI NIH HHS P30H027748 / PHS HHS EYO1792 / NEI NIH HHS
Language: English
Date published: 10/1991
Academic Unit: Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medical Genetics and Genomics; Ophthalmology and Visual Sciences
Record Identifier: 9983980082902771

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