English  |  正體中文  |  简体中文  |  Items with full text/Total items : 27248/39091
Visitors : 2415988      Online Users : 71
RC Version 4.0 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
Scope Adv. Search
LoginUploadHelpAboutAdminister

Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/10502

Title: 超氧歧化酵素、免疫調節蛋白之製備與其燙過癒合或抗潰瘍、以及免疫調節功能之探討
Preparation of Tat-Superoxide Dismutase and Tat-Fungus Immunomodulatory Protein and Their Possible Uses as Burn or Ulcer Healing and Immunomodulating Agents
Authors: 林棋財
Contributors: NTOU:Department of Aquaculture
國立臺灣海洋大學:水產養殖學系
Date: 2002-01
Issue Date: 2011-06-28T07:45:15Z
Publisher: 行政院農業委員會
Abstract: 摘要:超氧歧化酵素(SOD)它的功能是排除O2-,有報告指出SOD應用在創傷傷口能加速癒合和去除斑點及除皺.又SOD能調節神經細胞產生IL-1b、NO和TNF-a之功能,故SOD是維持身體健康扮演重要角色.食用菇近年來的研究發現具有增進免疫力與抗腫瘤,尤其真菌免疫調節蛋白(Fip)是一種存在食菇中分子量約13 kDa,具有活化T淋巴球、促進其細胞分裂以及IL-2、IFN-g和TNF-a等細胞激素的產生.目前Fip分別來自靈芝、金針菇、草菇與松杉靈芝.其蛋白質同質性達60%. SOD和Fip因來源不同其蛋白質的穩定性和活性不同,本實驗室對SOD的研究已有一段時間,發現由植物cDNA表現的SOD其穩定性和活性遠比動物高,其中有二種Cu/Zn-SOD和一種Mn-SOD非常安定.樟芝又名牛樟芝,對肝腫瘤及子宮頸腫瘤的治療非常有效,因只能在台灣牛樟樹生長.因樟芝是菇類唯一能代謝牛樟樹中大量抑菌性黃樟素而能生長的菇類.故本計畫擬以新鮮樟芝為材料,抽取RNA,合成cDNA或抽其染色體DNA,依據其他來源之SOD、Fip最保守區設計引子,用樟芝cDNA或染色體DNA為模板,以PCR增生DNA並進行篩選,應可篩得樟芝SOD、Fip之片段DNA,再進行5¢RACE和3¢RACE,最後得到全長SOD、Fip之DNA,繼之與表現型載体連接,送入微生物進行大量生產並純化,冀望找到的SOD比目前本實驗室已有的還安定,否則將採用植物SOD繼續進行以下實驗.SOD與Fip分子量介於32-13 KD之間,如何進入真核細胞是一大考驗,因此擬在N端分別接上9個銨酸之之Tat,則Tat-SOD, -Fip將容易進入細胞以發揮其功能.純化得到之Tat-SOD, -Fip將進行細胞免疫調節功能之測試,是否改變細胞之IL-1b、IL-2、NO和TNF-a等.又將與榮總整形外科馬旭副教授合作,進行燙傷癒合實驗,所得如為正面效果,將申請專利並與自然美公司合作應用於去除斑點及除皺的化妝品,或與順天堂合作添加在燙傷藥膏中.另將與葡萄王公司合作,將Tat-SOD, -Fip加在乳酸製品之類的飲料,或許能抗胃(十二指腸)潰傷飲料.第一年的工作著重於樟芝SOD、Fip之選殖與表現.第二年將進行SOD、Fip與Tat之連接,並製備較大量的Tat-SOD, -Fip.第三年進行細胞免疫調節功能之測試與燙傷癒合實驗.
Abstract:The role of superoxide dismutase (SOD) is to catalyze the dismutation of the superoxide ion (O2-) to hydrogen peroxide and molecular oxygen during oxidative energy processes. The reaction diminishes the destructive oxidative processes in cells. SOD was used as burn or ulcer healing, the protection of the skin against inflammatory reaction, periodontal use, the modulation of NO and cytokines. Thus, SOD plays very important role in maintaining healthy. Recent studies showed that medicinal mushrooms have physiological activities, especially the stimulation of the immune system and the prevention effects on cancer. They are reported having the capabilities to stimulate the proliferation of immune cells, increase the production of cytokines and tumor necrosis factor-γ. Their effects are believed to contribute to their physiological functions. In addition, evidences have showed that polysaccharides, polysaccharide-protein complexes and immunomodulatory proteins are known as the physiologically active components in mushrooms. Fungi immunomodulatory proteins (Fips) are one group of mushroom protein. The four cloned Fips show high homology in amino acid sequence to each other. Fips providing various activities on regulating immune system are the physiologically active components with identified structures in medicinal mushrooms. Antrodia camphorata, one kind of the medicinal mushrooms, can naturally parasites in the punk of Cinnamomum kanehirai Hay that is a special camphor tree locally existing in Taiwan. Therefore, this local mushroom shows a special meaning in Taiwan. In ethnologic medicine, A. camphorata has the activities of detoxification, anticancer and curing liver-complaint. This motivate us to clone full-length DNA of SOD and Fip from A. camphorata, then are introduced into expression vectors, we hope to obtain large amount of high activity and stable enzymes. The molecular mass of SOD and Fip was estimated to be 32-13 kDa, is not easily transferred into mammalian cell, if fused with Tat on N-terminus of both proteins, the barrier will be solved. First year, both full-length DNAs will be cloned. Second year, we will prepare large amount of Tat-SOD and Tat-Fip. Third year, we will perform burn or ulcer healing tests and immunomodulation in cells.
Relation: 91農科-3.1.3-糧-Z3(6)
URI: http://ntour.ntou.edu.tw/ir/handle/987654321/10502
Appears in Collections:[水產養殖學系] 研究計畫

Files in This Item:

There are no files associated with this item.



All items in NTOUR are protected by copyright, with all rights reserved.

 


著作權政策宣告: 本網站之內容為國立臺灣海洋大學所收錄之機構典藏,無償提供學術研究與公眾教育等公益性使用,請合理使用本網站之內容,以尊重著作權人之權益。
網站維護: 海大圖資處 圖書系統組
DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - Feedback