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
Externally publishedYes

Fingerprint

Aldehyde Dehydrogenase
Nucleotides
Polymorphism
Aldehydes
Single Nucleotide Polymorphism
Genes
Biotin
Streptavidin
Oxidoreductases
Fluorescence microscopy
Fluorescence Microscopy
Amines
Bacteria
DNA
Enzymes
Fluorescence
Alleles
Genotype
Polymerase Chain Reaction

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Electrochemistry

Cite this

Single nucleotide polymorphism genotyping of aldehyde dehydrogenase 2 gene using a single bacterial magnetic particle. / Yoshino, Tomoko; Tanaka, Tsuyoshi; Takeyama, Haruko; Matsunaga, Tadashi.

In: Biosensors and Bioelectronics, Vol. 18, No. 5-6, 01.05.2003, p. 661-666.

Research output: Contribution to journalArticle

@article{4518defdbe0c42ab845460e948a1123e,
title = "Single nucleotide polymorphism genotyping of aldehyde dehydrogenase 2 gene using a single bacterial magnetic particle",
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.",
keywords = "Aldehyde dehydrogenase 2, Bacterial magnetic particle (BMP), Magnetic bacteria, Single nucleotide polymorphism (SNP)",
author = "Tomoko Yoshino and Tsuyoshi Tanaka and Haruko Takeyama and Tadashi Matsunaga",
year = "2003",
month = "5",
day = "1",
doi = "10.1016/S0956-5663(03)00018-6",
language = "English",
volume = "18",
pages = "661--666",
journal = "Biosensors",
issn = "0956-5663",
publisher = "Elsevier Limited",
number = "5-6",

}

TY - JOUR

T1 - Single nucleotide polymorphism genotyping of aldehyde dehydrogenase 2 gene using a single bacterial magnetic particle

AU - Yoshino, Tomoko

AU - Tanaka, Tsuyoshi

AU - Takeyama, Haruko

AU - Matsunaga, Tadashi

PY - 2003/5/1

Y1 - 2003/5/1

N2 - 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.

AB - 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.

KW - Aldehyde dehydrogenase 2

KW - Bacterial magnetic particle (BMP)

KW - Magnetic bacteria

KW - Single nucleotide polymorphism (SNP)

UR - http://www.scopus.com/inward/record.url?scp=0037403577&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037403577&partnerID=8YFLogxK

U2 - 10.1016/S0956-5663(03)00018-6

DO - 10.1016/S0956-5663(03)00018-6

M3 - Article

VL - 18

SP - 661

EP - 666

JO - Biosensors

JF - Biosensors

SN - 0956-5663

IS - 5-6

ER -