Single nucleotide polymorphism genotyping of aldehyde dehydrogenase 2 gene using a single bacterial magnetic particle

Tomoko Yoshino, Tsuyoshi Tanaka, Haruko Takeyama, Tadashi Matsunaga

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

A single nucleotide polymorphism (SNP) genotyping for aldehyde dehydrogenase 2 gene (ALDH2) has been developed by using a nano-sized magnetic particle, which was synthesized intracellularly by magnetic bacteria. Streptavidin-immobilized on bacterial magnetic particles (BMPs) were prepared using biotin labeled cross-linkers reacting with the amine group on BMPs. ALDH2 fragments from genomic DNA were amplified using a TRITC labeled primer and biotin labeled primer pair, and conjugated onto BMP surface by biotin-streptavidin interaction. PCR product-BMP complex was observed at a single particle level by fluorescence microscopy. These complexes were treated with restriction enzyme, specifically digesting the wild-type sequence of ALDH2 (normal allele of ALDH2). The homozygous (ALDH2*1/*1), heterozygous (ALDH2*1/*2), and mutant (ALDH2*2/*2) genotypes were discriminated by three fluorescence patterns of each particle. SNP genotyping of ALDH2 has been successfully achieved at a single particle level using BMP.

Original languageEnglish
Pages (from-to)661-666
Number of pages6
JournalBiosensors and Bioelectronics
Volume18
Issue number5-6
DOIs
Publication statusPublished - 2003 May 1

Keywords

  • Aldehyde dehydrogenase 2
  • Bacterial magnetic particle (BMP)
  • Magnetic bacteria
  • Single nucleotide polymorphism (SNP)

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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