SNP genotyping in the beta(2)-adrenergic receptor by electronic microchip assay, DHPLC, and direct sequencing.
The beta(2)-adrenergic receptor (beta2AR) is the key target for the beta(2)-agonist drugs used for bronchodilation in asthma and chronic obstructive pulmonary disease. To detect four SNPs with amino acid variations at positions -47T/C (CysBUP19Arg), 46A/G (Gly16Arg), 79C/G (Gln27Glu), and 491C/T (Thr164Ile) in the beta 2AR gene, we used the electronic microchip assay, denaturing high-performance liquid chromatography (DHPLC), and direct sequencing. Genomic DNA samples were obtained from the blood of 84 Japanese healthy volunteers. The agreement rates of the first data set with the final data (allele calls) were 99.7% (332/333), 99.2% (246/248), and 96.7% (329/340). The percentages of no allele designation (ND) were 2.06% (7/340), 2.75% (7/255), and 0.00% (0/340) for the electronic microchip assay, DHPLC, and direct sequencing, respectively. Furthermore, we found three samples that had a novel haplotype.
Yoshida
Nobuyo
N
Department of Molecular, Cell Pharmacology, National Research Institute for Child Health and Development, 3-35-31 Taishido, Setagaya-ku, Tokyo 154-8567, Japan.
Nishimaki
Yuko
Y
Sugiyama
Masahide
M
Abe
Takashi
T
Tatsumi
Taiga
T
Tanoue
Akito
A
Hirasawa
Akira
A
Tsujimoto
Gozoh
G
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000097:blood / Q000737:chemistry* / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D007564:Japan / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics*; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018343:Receptors, Adrenergic, beta-2; D004247:DNA