| Co-Investigator(Kenkyū-buntansha) |
TABATA Osamu Kyoto University, Grad. School of Eng., Professor, 大学院・工学研究科, 教授 (20288624)
MASATO Taoka Tokyo Metropolitan University, Fac. Sci., Research Associate, 大学院・理学研究科, 助手 (60271160)
SHINOHARA Yasuo Inst. Genome Res., Professor, 教授 (60226157)
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| Research Abstract |
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
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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
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