A new method for detecting single nucleotide polymorphism using GFP-display.
The single nucleotide polymorphism (SNP) of aldehyde dehydrogenase-2 (ALDH2) codon 487, GAA (Glu) or AAA (Lys), was examined using green fluorescent protein (GFP)-display, an electrophoretic detection method for single amino acid changes. Although no shift in migration between the GFP-ALDH (Glu487) and GFP-ALDH (Lys487) fusion proteins was observed on SDS/urea gel, the two migrated to different positions when tagged with Asp. The SNP analysis was performed with GFP-ALDH-Asp3, and GFP-ALDH-Asp3 constructed from donors having the codon GAA/GAA, GAA/AAA or AAA/AAA was detected as different patterns as expected. GFP-display is potentially a unique method in SNP analysis, which does not require any special equipment or chemicals.
Aoki
Takashi
T
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido 061-0293, Japan. aokit@hoku-iryo-u.ac.jp
Satoh
Keiko
K
Imamura
Tomoko
T
Watabe
Hiroyuki
H
D000444:Aldehyde Dehydrogenase / Q000235:genetics; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D001224:Aspartic Acid / Q000737:chemistry; D003062:Codon; D004247:DNA / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D006801:Humans; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide
D003062:Codon; D010455:Peptides; D049452:Green Fluorescent Proteins; D001224:Aspartic Acid; D004247:DNA; C096127:ALDH2 protein, human; D000444:Aldehyde Dehydrogenase; D000071396:Aldehyde Dehydrogenase, Mitochondrial