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
N1 - Funding Information:
We would like to thank Dr Lawlor for suggestions to this manuscript. This work was funded in part by a Grant-in-Aid for Specially Promoted Research (2), No. 13002005 and Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.
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)
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U2 - 10.1016/S0956-5663(03)00018-6
DO - 10.1016/S0956-5663(03)00018-6
M3 - Article
C2 - 12706576
AN - SCOPUS:0037403577
SN - 0956-5663
VL - 18
SP - 661
EP - 666
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
IS - 5-6
ER -