Single nucleotide polymorphism genotyping of aldehyde dehydrogenase 2 gene using a single bacterial magnetic particle.
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.
Yoshino
Tomoko
T
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, 184-8588, Tokyo, Japan.
Tanaka
Tsuyoshi
T
Takeyama
Haruko
H
Matsunaga
Tadashi
T
D000444:Aldehyde Dehydrogenase / Q000737:chemistry / Q000235:genetics*; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D001426:Bacterial Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis / Q000737:chemistry; D020869:Gene Expression Profiling / Q000379:methods*; D005838:Genotype; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008280:Magnetics; D008565:Membrane Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods*; D008863:Microspheres; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D019809:Streptavidin / Q000737:chemistry
D001426:Bacterial Proteins; D020099:Coated Materials, Biocompatible; D008565:Membrane Proteins; C405425:MpsA protein, Magnetospirillum sp.; D019809:Streptavidin; C096127:ALDH2 protein, human; D000444:Aldehyde Dehydrogenase; D000071396:Aldehyde Dehydrogenase, Mitochondrial