Synthesis of Affinity Gels for Capillary Affinity Electrophoresis of Analysis of Nucleic Acids

用于核酸分析的毛细管亲和电泳亲和凝胶的合成

• 批准号: 06453150
• 负责人: AKASHI Mitsuru
• 金额: $ 4.74万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06453150/
• 关键词: Capillary Electrophoresis; Hydrogel; Affinity Ligad; Double Stranded DNA; Intercalator; Vinylmalachite Green; Polyvinyladenine; Capillary Coating; キャピラリー電気泳動; ポリビニルアルコール; 核酸; アフィニティー電気泳動; ポリビニールアデニン; 癌遺伝子; 配糖体ポリマー

High-resolution separation of nucleic acids (DNA and RNA) have been widely achieved using gel electrophoresis and HPLC.Recently, capillary gel electrophoresis (CGE) has been developing rapidly, as the resolving power of CGE is much higher than those of gel electrophoresis and HPLC.And the technique of affinity chromatography is also effective for analysis of nucleic acids. In this project, we investigated the capillary affinity gel electrophoresis (CAGE) of nucleic acids as development of an affinity mode of CGE.First, CAGE using polyacrylamide-poly (9-vinyladenine) conjugate gel was very effective for the recognition of thymine in nucleic acids and affinity separation of base-base mismatching DNA from original DNA (20mers) was achieved. Moreover, intercalator such as malachite green was immobilized in water soluble polymers to show the affinity to AT base pairs even in double stranded DNA.Moreover, we studied a new hydrogel for this system and capillary coating using water soluble polymers.

凝胶电泳和高效液相色谱已广泛应用于核酸（DNA和RNA）的高分辨率分离。近年来，毛细管凝胶电泳（CGE）因其分辨率远高于凝胶电泳和高效液相色谱而得到了迅速发展。亲和层析技术也适用于核酸的分析。在本项目中，我们研究了核酸的毛细管亲和凝胶电泳（CAGE）作为CGE亲和模式的发展。首先，使用聚丙烯酰胺-聚（9-乙烯基腺嘌呤）偶联凝胶的CAGE对胸腺嘧啶的核酸识别非常有效，并实现了碱基错配DNA与原始DNA （20mers）的亲和分离。此外，嵌入剂如孔雀石绿被固定在水溶性聚合物中，即使在双链DNA中也显示出对AT碱基对的亲和力。此外，我们还研究了一种用于该体系的新型水凝胶和采用水溶性聚合物的毛细管涂层。

项目成果

Mitsuru Akashi et al.: "Synthesis of Nonionic and Anionic Hydrogels Bearing a Monosaccharide Residue and Their Properties" Journal of Applied Polymer Science. 52. 1757-1763 (1994)

Mitsuru Akashi 等人：“带有单糖残基的非离子和阴离子水凝胶的合成及其特性”应用聚合物科学杂志。

明石 満 他: "光スイッチング機能を有するビニルマラカイトグリーンと二本鎖DNAとのその相互作用(2)" polymer preprints,Japan. 44. 2279-2280 (1995)

Mitsuru Akashi 等人：“具有光开关功能的乙烯基孔雀石绿及其与双链 DNA 的相互作用 (2)”聚合物预印本，日本 44. 2279-2280 (1995)。

Mitsuru Akashi, Akio Kishida et al.: ""Evaluation of the selective binding ability of oligodeoxynucleotides to poly (9-vinyladenine) using capillary affinity gel electrophoresis"" Analytical Science. Vol.10. 967-969 (1994)

Mitsuru Akashi、Akio Kishida 等人：“使用毛细管亲和凝胶电泳评估寡脱氧核苷酸与聚（9-乙烯基腺嘌呤）的选择性结合能力”“分析科学”。

Mitsuru Akashi, Akio Kishida et al.: ""The study of the interaction between vinyl malachite green having photo-switching ability and double stranded DNAs (2)"" Polymer Preprints, Japan. Vol.44. 2279-2280 (1995)

Mitsuru Akashi、Akio Kishida 等人：“具有光转换能力的乙烯基孔雀石绿与双链 DNA 之间相互作用的研究 (2)””聚合物预印本，日本。

Mitsuru Akashi et al.: "Evaluation of the Selective Binding Ability of Oligodeoxynucle otides to poly(9-vinyladenine)Using Capillary Affinity Gel Electrophoresis" Analytical Sciences. 10. 967-969 (1994)

Mitsuru Akashi 等人：“使用毛细管亲和凝胶电泳评估寡脱氧核苷酸与聚（9-乙烯基腺嘌呤）的选择性结合能力”分析科学。