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New mutant gene (transthyretin Arg 58) in cases with hereditary polyneuropathy detected by non-isotope method of single-strand conformation polymorphism analysis.
Second Department of Internal Medicine, Osaka University Medical School, Japan.
Single-strand conformation polymorphism (SSCP) was analyzed to detect a mutation in the transthyretin (TTR) gene from the mother and son showing polyneuropathy with carpal tunnel syndrome. DNA segments containing TTR coding sequence were amplified by polymerase chain reaction, heat denatured and electrophoresed on a neutral polyacrylamide gel. The single-stranded DNA fragments in the gel were transferred to a nylon membrane and hybridized with biotinylated TTR cDNA probe, followed with chemiluminescent DNA detection. The mobility shift was found in the fragments of exon 3 from the patients' DNA. Sequencing analyses of the exon 3 confirmed a T----G base change, resulting in a Leu 58----Arg substitution. TTR Arg 58 is the first mutant TTR gene that has been detected by SSCP analysis. The rapid and sensitive detection of new mutations at various sites on the TTR gene is hereafter possible by the present method in the facilities for non-radioactive experiments.
PMID: 1656975 [PubMed - indexed for MEDLINE]
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Cited by 2 PubMed Central articles
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Structure of Met30 variant of transthyretin and its amyloidogenic implications.
Terry CJ, Damas AM, Oliveira P, Saraiva MJ, Alves IL, Costa PP, Matias PM, Sakaki Y, Blake CC.
EMBO J. 1993 Feb; 12(2):735-41.
[EMBO J. 1993]
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'Cold SSCP': a simple, rapid and non-radioactive method for optimized single-strand conformation polymorphism analyses.
Hongyo T, Buzard GS, Calvert RJ, Weghorst CM.
Nucleic Acids Res. 1993 Aug 11; 21(16):3637-42.
[Nucleic Acids Res. 1993]