アフィニティーキャピラリー電気泳動による遺伝子診断


簡易かつ信頼性の高い一塩基変異検出方法を目指すアプローチの一つとして、 DNA-ポリアクリルアミドコンジュゲートを用いたアフィニティーキャピラリー電気泳動法 (ACE法)の開発に取り組んでいます。この方法は、 DNA二本鎖形成の親和性の差を利用する配列特異的な分離法です。本法を用いて癌遺伝子であるK-rasの正常体とコドン12の一塩基変異体を分離することに成功しました。

 

K-ras遺伝子コドン12のアフィニティキャピラリー電気泳動法による一塩基変異検出

K-ras遺伝子コドン12のアフィニティキャピラリー電気泳動法による一塩基変異検出

 

文献

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Affinity capillary electrophoretic DNA separation using PEG-oligodeoxyribonucleotide block copolymers: Relationship between peak resolution and affinity strength: Naoki Kanayama, Tohru Takarada, Ayumi Kimura, Hideaki Shibata, Mizuo Maeda, J. Sep. Sci., 31, 837-844, (2008).

(2)

Poly(ethylene glycol)-oligodeoxyribonucleotide block copolymers for affinity capillary electrophoretic separation of single-stranded DNAs with a single-base difference: Naoki Kanayama, Tohru Takarada, Ayumi Kimura, Hideaki Shibata, Mizuo Maeda, Reactive & Functional Polymers, 67, 1373-1380 (2007).

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Affinity capillary electrophoresis of DNA for detection of single-nucleotide  polymorphisms and point mutations: Comprehensive study for optimization of the weak affinity, Kae Sato, Miwa Onoguchi, Kazuo Hosokawa, and Mizuo Maeda, J. Chromatogr. A 1111, 120-126, (2006).

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Detection of single-base mutation by affinity capillary electrophoresis using a DNA-polyacrylamide conjugate, Kae Sato, Akira Inoue, Kazuo Hosokawa, Mizuo Maeda, Electrophoresis 26, 3076-3080 (2005).

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Capillary electrophoretic discrimination of single nucleotide polymorphisms using oligodeoxyribonucleotide-polyacrylamide conjugate as a pseudo-immobilized affinity ligand: optimum ligand length predicted by melting temperature values; Keiko Mishima, Tohru Takarada, Mizuo Maeda, Anal. Sci. 21, 25-29 (2005).

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Oligodeoxynucleotide-Modified Capillary for Electrophoretic Separation of Single-Stranded DNAs with a Single-Stranded DNAs with a Single-Base Difference; Takahisa Anada, Masako Ogawa, Hisashi Yokomizo, Yoshihisa Ozaki, Tohru Takarada, Yoshiki Katayama, Mizuo Maeda, Anal. Sci., 29, 73-77 (2003).

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The separation of oligodeoxynucleotides having a single-base difference by affinity capillary electrophoresis using oligodeoxynucleotide-polyacrylamide conjugate; Takahisa Anada, Takumi Arisawa, Yoshihisa Ozaki, Tohru Takarada, Yoshiki Katayama, Mizuo Maeda, Electrophoresis, 23, 2267-2273 (2002).


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