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

Biosens Bioelectron. 2003 May;18(5-6):661-6. doi: 10.1016/s0956-5663(03)00018-6.

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Dehydrogenase / chemistry
  • Aldehyde Dehydrogenase / genetics*
  • Aldehyde Dehydrogenase, Mitochondrial
  • Bacterial Proteins / analysis*
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / chemistry
  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • Coated Materials, Biocompatible / chemical synthesis
  • Coated Materials, Biocompatible / chemistry
  • Gene Expression Profiling / methods*
  • Genotype
  • Humans
  • Immunoassay / methods*
  • Magnetics
  • Membrane Proteins / analysis*
  • Membrane Proteins / biosynthesis
  • Membrane Proteins / chemistry
  • Microscopy, Fluorescence / methods*
  • Microspheres
  • Polymorphism, Single Nucleotide / genetics*
  • Streptavidin / chemistry

Substances

  • Bacterial Proteins
  • Coated Materials, Biocompatible
  • Membrane Proteins
  • MpsA protein, Magnetospirillum sp.
  • Streptavidin
  • ALDH2 protein, human
  • Aldehyde Dehydrogenase
  • Aldehyde Dehydrogenase, Mitochondrial