A new method for detecting single nucleotide polymorphism using GFP-display.

Takashi Aoki; Keiko Satoh; Tomoko Imamura; Hiroyuki Watabe

doi:10.1016/j.jbbm.2004.04.019

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A new method for detecting single nucleotide polymorphism using GFP-display.

Takashi Aoki·Keiko Satoh·Tomoko Imamura·Hiroyuki Watabe

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Abstract

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.

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Aldehyde Dehydrogenase Aldehyde Dehydrogenase, Mitochondrial Aspartic Acid Codon DNA Electrophoresis, Polyacrylamide Gel Escherichia coli Genetic Techniques Green Fluorescent Proteins Humans Peptides Plasmids Polymorphism, Single Nucleotide Journal Article Research Support, Non-U.S. Gov't

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Takashi Aoki, Keiko Satoh, Tomoko Imamura, Hiroyuki Watabe. 2004-07-30. A new method for detecting single nucleotide polymorphism using GFP-display.. https://doi.org/10.1016/j.jbbm.2004.04.019

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