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研究生:莊政益
研究生(外文):Cheng- Yi
論文名稱:成人血清中抗化膿性鏈球菌57 kDa膜蛋白之鑑定
論文名稱(外文):Identification of a 57 kDa membrane protein for group A Streptococcus
指導教授:陳凌雲陳凌雲引用關係
指導教授(外文):Ling-Yun Chen
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生化暨生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:50
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Streptococcus pyogenes(化膿性鏈球菌)為格蘭氏陽性致病菌又稱為group A streptococcus,GAS(A群鏈球菌),透過對人類皮膚及喉嚨的侵襲會造成如咽喉炎、猩紅熱等,且容易併發嚴重的後遺症如壞死性筋膜炎(necrotizing fasciitis)、敗血症(sepsis)和鏈球菌毒性休克症候群(streptococcal toxic shock syndrome)。GAS 透過表達許多的毒性因子如 M protein、C5a peptidase 而達到對宿主侵襲、感染和逃避免疫反應的能力。根據過去淑玲學姐的實驗中發現,成人血清會認識一個 GAS 57 kDa 的蛋白,且兒童血清對這個蛋白的辨識能力較低,這樣的結果再比較 CDC 猩紅熱病例的年齡分佈似乎有著相關性。但是過去利用各種膜蛋白萃取方式並不能有效的達到萃取的目的,即使利用陰離子親合性管柱DE-52、High Q 或疏水性離子管柱 phenyl column 亦無法有效的得到純化且穩定性高的蛋白。在本論文的研究中,我們試著以另一個策略來萃取與純化這個蛋白,在結果中以加熱的動作得到這個 GAS 57 kDa 蛋白的萃取液,更以陰離子親合性管柱 High Q 得到大量且純化的蛋白,再經過 MALDI-TOF 鑑定後發現此一特定蛋白為 M protein serotype 12 precursor。依據這個結果將有助於我們繼續研究可應用在 GAS 早期診斷上的工具。

Streptococcus pyogenes, also called group A streptococcus (GAS), is a Gram-positive bacterium. GAS can cause infectious diseases in skin or throat of human, including uncomplicated pharygitis and scarlet fever, and develop into severe complications , including necrotizing fasciitis, sepsis and streptococcal toxic shock syndrome. GAS can induce virulence factor expression such as M protein and C5a peptidase and increase invation、infection and immune evasion of host cells and tissue.In our previous study, we found hat M.W.57kDa protein of GAS can be recognized by adult serum but poorly recognized by children serum (the <14 years old serum). This result is association with distribution of age from CDC report. In our previous study,it is not effective to purify various membrane protein with anion affinity chromatography column DE-52、High Q and hydrophobic interaction phenyl column.In this study,we exploit a new strategy to searching for method of extraction and purification for M.W.57kDa protein of GAS. At first,we get extraction solution of M.W.57kDa protein of GAS by using heating.The next, we get a great quantity of purified protein by using anion affinity chromatography column High Q. At last, we found this protein is M protein serotype 12 precursor by usig MALDI-TOF identify. This finding will help us further studying in the future and can be used as tool of early diagnosis.

目錄
圖次……………………………………2
縮寫表…………………………………3
中文摘要………………………………4
英文摘要………………………………5
前言……………………………………6
研究動機………………………………18
實驗材料與方法………………………20
結果……………………………………25
討論……………………………………41
參考文獻………………………………45
附錄……………………………………48


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