Rapid and efficient molecular analysis of gyrate atrophy using denaturing gradient gel electrophoresis

Y. Mashima, T. Shiono, George Inana

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Purpose. A generalized biochemical deficiency of the mitochondrial matrix enzyme ornithine aminotransferase (OAT) is the inborn error in gyrate atrophy (GA), an autosomal recessive blinding disease of the retina and choroid of the eye. Because mutations in the OAT gene show a high degree of molecular heterogeneity in GA, the authors set out to determine the mutations by rapid and efficient methods. Methods. The mutations in the OAT gene were determined by a combination of polymerase chain reaction (PCR) amplification of gene sequences, analysis by denaturing gradient gel electrophoresis (DGGE), and direct DNA sequencing. Results. Eleven different mutations in 21 (95.5%) out of 22 mutant OAT alleles from 11 patients were identified: six missense mutations, three nonsense mutations, one 2 bp-deletion, and one splice acceptor mutation. A silent polymorphism of Asn (AAC)378 to Asn (AAT) was also observed. Conclusions. The combination of PCR amplification of the gene sequences, DGGE analysis, and direct sequencing is a rapid and efficient method for detection of mutations in GA cases. The diversity of the mutations attests to the enormous genetic heterogeneity in this disease.

Original languageEnglish
Pages (from-to)1065-1070
Number of pages6
JournalInvestigative Ophthalmology and Visual Science
Volume35
Issue number3
StatePublished - Jan 1 1994

Fingerprint

Gyrate Atrophy
Denaturing Gradient Gel Electrophoresis
Ornithine-Oxo-Acid Transaminase
Mutation
Gene Amplification
Choroid Diseases
Polymerase Chain Reaction
Genetic Heterogeneity
Nonsense Codon
Missense Mutation
DNA Sequence Analysis
Genes
Sequence Analysis
Retina
Alleles

Keywords

  • denaturing gradient gel electrophoresis
  • genetic heterogeneity
  • gyrate atrophy
  • ornithine aminotransferase
  • polymerase chain reaction

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Rapid and efficient molecular analysis of gyrate atrophy using denaturing gradient gel electrophoresis. / Mashima, Y.; Shiono, T.; Inana, George.

In: Investigative Ophthalmology and Visual Science, Vol. 35, No. 3, 01.01.1994, p. 1065-1070.

Research output: Contribution to journalArticle

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