綠竹筍生長過程蛋白質體變化及其抗體庫之建立

Study on the Proteome of the Fast Growing Bamboo Shoots and the Construction of its Antibody Bank

吳裕仁(Yu-Jen Wu)

指導教授 ： 莊榮輝

國立臺灣大學/生命科學院/微生物與生化學研究所/博士(2006年)

https://doi.org/10.6342/NTU.2006.00266

摘要

針對竹筍快速生長的特性，本計畫利用『蛋白質體』工具，比對生長過程中蛋白質的總體變化。竹筍由未出土到生長六十公分的過程中，水溶性蛋白質具有消長的變化，目前鑑定及分析消長色點的身分，並且配合酵素活性分析，發現綠竹筍快速生長時，細胞壁多醣類快速轉變，糖解作用以及檸檬酸循環作用加速，以提供竹筍生長所需的能量；五碳醣磷酸路徑表現量保持恆定，持續提供核醣及NADPH，以提供綠竹生長時核酸、胺基酸及固醇類合成所需。另外一些參與生長的G-protein, elongation factor等蛋白質，以及防禦機制的蛋白質及代謝酵素表現量都增多，可能也是造成竹筍快速生長的原因。
另外以竹筍全體蛋白質進行單株抗體庫製備的研究，以全部的蛋白質進行小鼠免疫，再利用階段式抗體篩選方法，並且改變抗體製備及篩選流程，目前已經成功選出約192株不同單株抗體，此種方法是一全新的作法及概念，可以省去抗原的純化步驟，並且在短時間內製備出多樣化的單株抗體。而且結合蛋白質體及抗體庫的平台，是一個非常有用基礎技術平台，可對任何生物系統做相似的應用，產生任何蛋白質體的抗體庫 (小白鼠系統除外)。得到抗體庫後接著可進行抗體晶片的製作，以供相關生物系統在其細胞學、生理學、病理學等各範疇的基礎與應用研發。

關鍵字

綠竹筍 ； 蛋白質體 ； 抗體庫

並列摘要

This project employing the proteomics method to investigate the overall changes of proteins during the growing process in bamboo which is fast-growing in nature. It was observed that the levels of water-soluble proteins fluctuate in the growing stage from underground to 60 cm-shoot high. Identifying the spots which undergo changes during the growing process in combination with enzyme activity analysis, it were discovered that polysaccharides of cell walls change rapidly, glycolysis and tricarboxylic acid cycle metabolism accelerate to generate the energy for growing, the level of pentose phosphate pathway remains stable to supply ribose and NADPH for the synthesis of nucleic acids, amino acids, and steroids. Additionally, the amounts of some proteins related to growing such as G-protein and elongation factor, proteins with defense mechanism and metabolism enzymes increase to cause the fast-growing of bamboo.
In a separate research, the establishment of monoclonal antibody bank using all of the proteins of bamboo at once to perform immune test on mice was investigated. So far 192 monoclonal antibodies have been isolated using the method of stage antibody screening with modifications on the antibody preparation and screening process. This method is a new concept and process. The purification process of antigen could be eliminated and many multiple monoclonal antibodies can be prepared in a short time.
It is a platform combining proteomics and antibody bank. Many applications based on this platform can be performed. It can be applied to any biological system to generate antibody of any protein (except mice). Once the antibody bank is established, the making of the antibody chips can be conducted to provide research and development tools for cell biology, physiology, pathology and other related fields。

並列關鍵字

Bambusa oldhamii ； proteome ； antibody bank

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