A new tool for DNA research : Slalom chromatography

DNA 研究的新工具：回转色谱

• 批准号: 03559006
• 负责人: KASAI Ken-ichi
• 金额: $ 4.22万
• 依托单位: Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03559006/
• 关键词: DNA; size separation; slalom chromatography; HPLC; nucleic acid; スラロ-ムクロマトグラフィ-; 環状DNA; ス-パ-コイル; リラックス; 付着末端; λ-ファ-ジ

Slalom chromatography is a new and unique procedure for size fractionation of large DNA molecules, which was invented by our group. In contrast to all traditional chromatographic modes that are based on equilibrium phenomena, slalom chromatography is based on a hydrodynamic phenomenon, which was found for the first time. By the present research project, extremely useful results were obtained from viewpoints of not only theoretical aspects but also future applications. Principal factors which affect size separation of DNA molecules proved to be size of packing particle, flow rate and viscosity of eluents, and temperature. This strongly suggests that the longer a DNA molecule, the more strong frictional force is generated because of laminar flow. Behavior of circular form DNA molecules was different for linear molecules and found to depend on their topological differences, that is, super coil, relaxd and single strand. Therefore, slalom chromatography seems to be very useful for not only size separation but also physicochemical studies of DNA.In order to widen the dynamic range, various attempts have been made. Resolution of smaller DNA molecules was slightly improved by the combination of slalom mode and weak hydrophobic mode, but it was difficult to improve resolution of larger molecules. Since utility of size fractionation of cDNAs was also demonstrated, it should contribute to the human genom project.

回转色谱法是本课题组发明的一种新颖独特的DNA大分子分离方法。与所有基于平衡现象的传统色谱模式相反，回转色谱基于流体动力学现象，这是第一次发现。通过本研究项目，不仅从理论方面而且从未来应用的角度获得了非常有用的结果。影响DNA分子大小分离的主要因素是填充颗粒的大小、洗脱液的流速和粘度以及温度。这有力地表明，DNA分子越长，由于层流而产生的摩擦力就越强。环状DNA分子的行为与线性分子不同，并发现这取决于它们的拓扑结构差异，即超螺旋、松弛和单链。因此，回旋色谱法不仅对DNA的大小分离，而且对DNA的物理化学研究都是非常有用的。结合回旋模式和弱疏水模式对小分子DNA的分辨率略有提高，但对大分子DNA的分辨率提高困难。由于cDNA的大小分级的效用也被证明，它应该有助于人类基因组计划。

项目成果

Kasai, K.: "Intelligent research tools in bioscience : Affinity technique and others." Bunseki. 204. 968-974 (1991)

Kasai, K.：“生物科学中的智能研究工具：亲和技术等。”

Kasai, K.: "Size-dependent chromatographic separation of nucleic acids." J.Chromatogr.618. 203-221 (1993)

Kasai, K.：“核酸的尺寸依赖性色谱分离。”

Ngo,T.,(Editor): "Molecular Interaction in Bioseparations" Plenum Publishing Co,,(in press),

Ngo,T.,（编辑）：“生物分离中的分子相互作用”Plenum Publishing Co,,（印刷中）,

笠井献一: "バイオ時代の賢い研究法;アフィニティー技術を中心に" ぶんせき. 204. 968-974 (1991)

Kenichi Kasai：“生物时代的智能研究方法；关注亲和技术”Bunseki 204. 968-974 (1991)。

Shimura,K.: "Affinophoresis:Selective electrophoretic separation of proteins by using a specific carrier.“Methods in Enzymology"(Karger,B.L.,Hancock,W.S.,eds)" Academic Press,New York,

Shimura, K.：“亲和电泳：使用特定载体对蛋白质进行选择性电泳分离”（Karger，B.L.，Hancock，W.S.，编辑）”学术出版社，纽约，