Improvement of DNA chip system with probe-on-carriers for practical uses

载体上探针DNA芯片系统的改进以实用化

• 批准号: 18310135
• 负责人: TSUKAHARA Toshifumi
• 金额: $ 11.11万
• 依托单位: Japan Advanced Institute of Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18310135/
• 关键词: DNA Chin; Microarrav; Porous Glass; Synthesis of Nucleic Acids; Probe-on-Carrier; Genotype Analysis; Gene Expression Analysis; Order-Maid Medicine; 超分子化学; 遺伝子; ゲノム; バイオ関連機器; 固相担体

In this study, we are improving a method to generate DNA chips with "probe-on-carriers", immobilized oligonucleotide probes on the solid phase, for practical uses. In this DNA chip, each oligonucleotide was synthesized on porous glass as a solid phase-carrier, and can be used as an immobilized probe for the complementary target sequence. Therefore, high-quality DNA chips can be fabricated easily and economically.First, we optimized synthesis of DNA probes on porous glass resins by using a new hydrophobic linker and a new non-aqueous reagent for the deprotection process. We also proposed a new strategy called "Protected DNA Probes (PDP) method" in which appropriately protected bases can bind highly selectively to the complementary bases even without removal of their base protecting groups. This PDP strategy could guarantee highly efficient synthesis of DNA probes on controlled porous glass with high purity and thereby could eliminate the time-consuming procedures for isolation of DNA pr … More obes. SNPs were successfully recognized each other by using PDPs immobilized on glass plates, suggesting its potential usefulness. Moreover, we succeeded to develop new method for the SNP analysis by using a chemical or photochemical ligation technique on plates with high coupling efficiency. These methods showed markedly high match/mismatch discrimination ability. Therefore, these technologies resulted in significant improvement of the base discrimination ability in DNA chip system with probe-on-carriers for practical uses.We also tried to develop DNA microarrays with probe-on-carriers for gene expression analysis. The porous glass supports gave an excellent result for a 120 mer-long oligonucleotide synthesis. The yield was almost 20 times as much as by using cross-linked polystyrene, the most popular support for oligonucleotide synthesis. Therefore, by using probe-on-carriers with different probe lengths, an economical device, which could analyze not only SNPs but also gene expression at one time, could be developed. Less

在这项研究中，我们正在改善一种用“探针载体”生成DNA芯片的方法，用于实用用途。在此DNA芯片中，将每种寡核苷酸合成在多孔玻璃上作为固相载体，并可以用作完整目标序列的固定探针。因此，高质量的DNA芯片可以轻松，经济上的制造。首先，我们通过使用新的疏水性接头和新的非水剂进行脱废过程，优化了多孔玻璃树脂上DNA问题的合成。我们还提出了一种称为“受保护的DNA探针（PDP）方法”的新策略，其中适当保护的基础即使没有去除其碱基保护组也可以高度选择性地与完整碱基结合。这种PDP策略可以保证具有高纯度的受控多孔玻璃上DNA问题的高效合成，从而可以消除隔离的dna pr…更多物体的耗时程序。通过使用固定在玻璃板上的PDPS成功识别SNP，表明其潜在的有用性。此外，我们成功地通过使用高耦合效率的板上使用化学或光化学连接技术开发了SNP分析的新方法。这些方法表现出明显高的匹配/不匹配歧视能力。因此，这些技术通过用于实际用途的探针载体来显着提高DNA芯片系统中的基础歧视能力。我们还试图使用用于基因表达分析的探针载体来开发DNA微阵列。多孔玻璃载体为120个Mer-long寡核苷酸合成带来了极好的结果。该产量是使用交联聚苯乙烯的20倍，这是对寡核苷酸合成的最流行的支持。因此，通过使用具有不同探针长度的探针载体，可以开发一种经济装置，不仅可以分析SNP，而且可以一次分析基因表达。较少的

项目成果

Okinaga T.,Tsukahara T., Tajima Y., Ozono K, Nishino I., Nonaka I., Toda T : Aberrant neuromuscular junctions and delayed terminal muscle fiber maturation in a -dystroglycanopathies

Okinaga T.，Tsukahara T.，Tajima Y.，Ozono K，Nishino I.，Nonaka I.，Toda T：α-肌营养不良症中的异常神经肌肉接头和延迟终末肌纤维成熟

• 发表时间: 2006
• 期刊: Hum. Mot Genet 15
• 作者: Taniguchi;M.;Kurahashi;H.;Noguchi;S.;Fukudome;T
• 通讯作者: T

Expression profiling of muscles from Fukuyama-type congenital muscular dystrophy and laminin-alpha2 deficient congenital muscular dystrophy; is congenital muscular dystrophy a primary fibrotic disease?

福山型先天性肌营养不良症和层粘连蛋白-α2缺陷型先天性肌营养不良症的肌肉表达谱；

• 发表时间: 2006
• 期刊: Biochem Biophys Res Commun. 342
• 作者: Taniguchi M;Kurahashi H;Noguchi S;Sese J;Okinaga T;Tsukahara T;Guicheney P;Ozono K;Nishino I;Morishita S;Toda T
• 通讯作者: Toda T

Development of a next generation oligo-DNA microarray with probe-on-carriers for SNPs analysis

开发用于 SNP 分析的带有载体探针的下一代寡 DNA 微阵列

• 发表时间: 2006
• 作者: Tsukahara T;Nagasawa H;Ohkubo A;Seio K;Sekine M and Fuke S
• 通讯作者: Sekine M and Fuke S

Alternative splicing variants of Mef2c in PI9 cells and P I9CL6

PI9 细胞和 P I9CL6 中 Mef2c 的选择性剪接变体

• 发表时间: 2007
• 作者: Ab;Hakim;N-H.;Tsukahara;T.;Suzuki;H
• 通讯作者: H

Analysis of regulation mechanism in alternative splicing by NSSR/SRp38

NSSR/SRp38选择性剪接调控机制分析

• 发表时间: 2007
• 作者: Konishi;T.;Tsukahara;T.;Suzuki;H
• 通讯作者: H