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研究生:王暐嘉
研究生(外文):WANG WEI-CHIA
論文名稱:大豆胜肽抑制腸病毒71型感染之研究
論文名稱(外文):Inhibitory Effect of Soybean Peptide on Enterovirus 71 Infection
指導教授:林雅菁林雅菁引用關係
指導教授(外文):LIN YA-CHING
口試委員:吳聲祺鄭鈞文
口試委員(外文):WU SHENG-CHICHENG CHUN-WEN
口試日期:2018-08-30
學位類別:碩士
校院名稱:輔英科技大學
系所名稱:生物科技系碩士班
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:2018
畢業學年度:107
語文別:中文
論文頁數:66
中文關鍵詞:腸病毒71型大豆胜肽抑制感染
外文關鍵詞:Enterovirus 71Soy peptideInhibition of infection
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腸病毒71型是一種重要的人類致病性病毒,其併發症包括腦膜炎、急性心肺衰竭、急性肢體麻痺症候群等。目前尚未開發出有效的疫苗及治療腸病毒71型的標準方法,因此,平時做好預防就成為最重要的課題。大豆是一種營養價值高又普及的豆類食品,其胜肽據研究顯示具有抗癌、降高血壓、抗糖尿病、降膽固醇、抗氧化、抗肥胖及免疫調節等功能。本研究利用高營養價值的大豆胜肽進行抑制腸病毒71型感染力之探討。在研究中,利用鹼性蛋白酶與米麴菌蛋白酶製備大豆胜肽,所獲得之胜肽液以單獨或混和的方式進行抑制腸病毒71型感染效用之測試。結果顯示,大豆胜肽具有抑制腸病毒71型感染之效用。
Enterovirus 71 is an important human pathogenic virus. Among hundreds of enteroviruses, Enterovirus 71 causes the most serious complications and deaths. Its complications include meningitis, acute cardiopulmonary failure, Acute limb paralysis syndrome. Currently there is no effective vaccine and standard treatment for enterovirus 71, therefore, prevention is usually the most important issue. Soybeans are high in nutritional value and popularity of legumes. Their peptides have been shown to have anti-cancer, antihypertensive, anti-diabetic, cholesterol-lowering, antioxidant, anti-obesity and immune regulation functions. In this study, the effect of high-nutrient soybean peptides on the inhibition of enterovirus 71 infectivity was investigated. The soybean peptide was prepared by using alkaline protease and rice bran protease. The obtained peptide solution on inhibiting enterovirus type 71 infection was examined either alone or in combination. Our results show that the soybean peptide displayed the inhibitorty effect on enterovirus 71 infection.


誌謝 i
中文摘要 ii
英文摘要 iii
圖目錄 vii
第一章 緒論 1
第二章 文獻探討 3
第一節 大豆與大豆胜肽胺基酸 3
第二節 生物活性胜肽 3
第三節 大豆蛋白的生物活性功用 5
I. 抗糖尿病 5
II. 降高血壓 5
III. 降膽固醇 6
IV. 抗氧化 7
V. 抗肥胖 8
VI. 免疫調節 10
VII. 抗癌 11
VIII. 抗病毒效用 13
第四節 腸病毒 14
I. 簡介 14
II. 腸病毒的特性 15
III. 腸病毒的傳播途徑及防治 16
第三章 研究方法 18
第一節 實驗流程圖 18
第二節 實驗材料 19
I. 實驗藥品 19
II. 實驗儀器 21
第三節 培養基製備 21
第四節 細胞培養 21
第五節 腸病毒71型之病毒放大及純化 22
第六節 病毒力價測試 23
第七節 大豆胜肽製備 24
第八節 利用蛋白質濃縮管分離胜肽大小 25
第九節 L-Leucine 標準檢量曲線製作 25
第十節 水解率測定步驟及計算 26
第十一節大豆蛋白吸附病毒後力價測試 28
第十二節FITC螢光染色 28
第十三節蛋白質分析 (SDS-PAGE) 29
第十四節病毒核酸萃取 31
第十五節反轉錄聚合酶鏈鎖反應 (Reverse transcriptase-polymerase chain reaction)之檢測 32
第十六節聚合酶鏈鎖反應 (Polymerase chain reaction) 32
第四章 結果與討論 33
第一節 利用VIVASPIN6分離各個胜肽液的分子大小 33
第二節 確認不同水解酶所水解出的胜肽液經0.45μm過濾的胜肽大小 33
第三節 利用L-Leucine 標準檢量曲線測定水解率 34
第四節 胜肽液抗腸病毒71型效用測試 34
I. 腸病毒力價103TCID50測試 34
II. 腸病毒力價104TCID50測試 35
第五節 胜肽液對於腸病毒71型RNA的效用測試 36
第六節 FITC螢光染色檢測感染現象 36
第五章 結論與建議 38
第六章 參考文獻 39

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