Analysis of mismatched DNA by mismatch binding ligand (MBL)-Sepharose affinity chromatography

被引:6
作者
Goto, Yuki
Suda, Hitoshi
Kobori, Akio
Nakatani, Kazuhiko
机构
[1] Osaka Univ, Inst Sci & Ind Res, SANKEN, Ibaraki 5670047, Japan
[2] Kyoto Univ, Grad Sch Engn, Dept Synthet Chem & Biol Chem, Kyoto 6158510, Japan
关键词
mismatch binding ligand; affinity chromatography; mutation detection; melting temperature; single nucleotide polymorphism;
D O I
10.1007/s00216-007-1323-y
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Mismatch binding molecules (MBLs), strongly and selectively bound to the mismatched base pair in duplex DNA, were immobilized on Sepharose. Three MBL-Sepharose columns were prepared with three MBLs, naphthyridine dimer (ND), naphthyridine-azaquinolone (NA), and aminonaphthyridine dimer (amND), which exhibited different binding profiles to the mismatched base pairs. These three MBL-Sepharose columns showed characteristic elution profiles for DNA duplexes containing mismatched base pairs. The ND-Sepharose column separated the G-G and G-A mismatched DNA from fully matched duplexes. The NA-Sepharose column separated the A-A and G-A mismatched DNA from other DNA duplexes. The amND-Sepharose column separated the C-C mismatched DNA. These chromatographic profiles were very consistent with the binding preference of each MBL. By changing the elution conditions from sodium hydroxide to sodium chloride, MBL-Sepharose columns were also able to separate the mismatched DNA that weakly bound to the MBL from fully matched DNA duplex.
引用
收藏
页码:1165 / 1173
页数:9
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