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

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.

• Authors: Y Mashima, T Shiono, G Inana
• Journal: Investigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 35 Issue 3 Pg. 1065-70 (Mar 1994) ISSN: 0146-0404 [Print] United States
• PMID: 8125717 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA Primers
• DNA
• Ornithine-Oxo-Acid Transaminase

Topics

• Alleles
• Base Sequence
• DNA (analysis)
• DNA Mutational Analysis
• DNA Primers
• Electrophoresis, Polyacrylamide Gel
• Gyrate Atrophy (enzymology)
• Humans
• Mitochondria (enzymology)
• Molecular Biology
• Molecular Sequence Data
• Mutation
• Ornithine-Oxo-Acid Transaminase (genetics)
• Polymerase Chain Reaction
• Polymorphism, Genetic
• Protein Denaturation