Development for Advanced Bio-nanodevice for Functional Genomics and Proteomics

用于功能基因组学和蛋白质组学的先进生物纳米器件的开发

• 批准号: 13124206
• 负责人: BABA Yoshinobu
• 金额: $ 30.85万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13124206/
• 关键词: Nanobio; Nanofabrication; Genome; Biotechnology; Proteome

Since human genome sequencing has been completed, human genome project will quickly move on to the post genome sequencing era, including single nucleotide polymorphism (SNP) analysis, functional genomics, mutation analysis, transcriptome analysis, proteome analysis, metabolome analysis, and systems biology. Capillary array electrophoresis with 96-384 capillaries plays a vital role in the genome sequencing era, but in the post Human Genome sequencing era, further development of analytical technology for DNA, mRNA, protein, and metabolites is highly required for future medicine and new drug discovery technology based on functional genomics and proteomics. We developed nan-biodevice for DNA and genomic analysis. DNA fragments up to 1000 bp is separated within 10-120 s by microchip technology, Y-chromosomal polymorphism is analyzed within 90s. Haplotyping from human blood sample was achieved within 2 min by microchip. Array of nano-pillar, which is 200 nm wide and 4000 nm tall, is successf … More ully applicable to fast separation of DNA within 10-25s. The nano-biodevice is extremely useful for fast separation of protein samples from several kDa to 200 kDa within 15 seconds. Utilizing this technique, complex protein mixture extracted from human T cell line, lymphoblastic Jurkat cells, were separated within 15s with high reproducibility. The target proteins of the Jurkat cells, which increase after heat-shock apoptosis, wore detected and identified within by this method, while several hours are required for conventional 2-DE analysis. This novel technique on a microchip will ofter the enormous advantages for high through-put screening systems of proteome analysis, especially for rapid detection of small amount of target proteins, those are idiotypic for a specific physiologic or pathologic state of cells or tissues. The nano-biodevice will be applicable to fast-analysis of sugar chain on the glycoproteins. We developed high-sensitive LED induced fluorescence detection system for nano-biodevice and applied to analysis of fluorescent labeled sugar chain. Complex mixture of sugar chains digested from glycoproteins (AGP and IgG) is separated and detected within 60 s with high-resolution. The nano-biodevice will be applicable to the SNP analysis of a single genomic DNA molecule. Less

随着人类基因组测序的完成，人类基因组计划将迅速进入后基因组测序时代，包括单核苷酸多态性（SNP）分析、功能基因组学、突变分析、转录组分析、蛋白质组分析、代谢组分析和系统生物学。96-384毛细管的毛细管阵列电泳在基因组测序时代起着至关重要的作用，但在后人类基因组测序时代，DNA、mRNA、蛋白质和代谢物的分析技术的进一步发展是基于功能基因组学和蛋白质组学的未来医学和新药发现技术的高度要求。我们开发了用于DNA和基因组分析的纳米生物装置。通过微芯片技术在10-120 s内分离高达1000 bp的DNA片段，在90 s内分析Y染色体多态性。通过微芯片在2分钟内实现了人血液样本的单倍型分型。成功制备出宽200 nm、高4000 nm的纳米柱阵列， ...更多信息 特别适用于10- 25秒内DNA的快速分离。纳米生物装置对于在15秒内快速分离从几kDa到200 kDa的蛋白质样品非常有用。利用该技术，从人T细胞系淋巴母细胞Jurkat细胞中提取的复杂蛋白质混合物在15秒内分离，具有高重复性。该方法可在短时间内检测到Jurkat细胞热休克凋亡后增加的靶蛋白，而常规的双向电泳分析需要数小时。这种新的微芯片技术将为蛋白质组分析的高通量筛选系统提供巨大的优势，特别是对于快速检测细胞或组织的特定生理或病理状态的少量目标蛋白。该纳米生物器件可应用于糖蛋白糖链的快速分析。研制了高灵敏度的纳米生物器件LED诱导荧光检测系统，并应用于荧光标记糖链的分析。从糖蛋白（AGP和IgG）中消化的糖链的复杂混合物在60秒内以高分辨率分离和检测。该纳米生物器件将适用于单个基因组DNA分子的SNP分析。少

项目成果

Mari Tabuchi: "The Separation Carrier for High-Speed Proteome Analysis by Capillary Electrophoresis"Electrophoresis. 22(16). 3449-3457 (2001)

Mari Tabuchi：“毛细管电泳高速蛋白质组分析的分离载体”电泳。

M.Hino: "Requirement of Continuous Transcription for Synthesis of Sufficient Amount of Protein by Cell Free Rapid Translation System"Protein Expression and Purification. 24. 255-259 (2002)

M.Hino：“通过无细胞快速翻译系统合成足够量蛋白质的连续转录的要求”蛋白质表达和纯化。

F.Xu: "Relationship between Width and Migration Time Used in Double-Stranded DNA Separation by Microchip Electrophoresis"Analyst. (印刷中). (2003)

F.Xu：“微芯片电泳分离双链 DNA 时的宽度与迁移时间的关系”分析员（2003 年出版）。

M.Taoka: "V-1, a protein expressed transiently during marine cerebellar development regulates actin polymerization via interaction with capping protein"J.Biol.Chem.. 278(8). 5864-5870 (2003)

M.Taoka：“V-1 是海洋小脑发育过程中短暂表达的一种蛋白质，通过与加帽蛋白相互作用调节肌动蛋白聚合”J.Biol.Chem.. 278(8)。

M.Tabuchi: "Nanospheres for DNA Separation Chips"Nature Biotechnology. 22. 337-340 (2004)

M.Tabuchi：“用于 DNA 分离芯片的纳米球”自然生物技术。