隨著越來越多的細菌基因體被全定序，透過搜尋資料庫的方法，可以發現有許多基因非常類似細菌素(bacteriocin)或跟細菌素有關的誘導物質(autoinducer peptide，縮寫為AIP)，參考細菌素的一些特性，如帶正電、分子量大小，是否具備GG-motif和序列相似性…等方法，在NCBI、KEGG、PFAM…等資料庫中，找到了七個有可能為細菌素的基因，分別是: SEQ 31、SEQ 34、SEQ 35、SEQ 36、SEQ 17、SEQ 29 and SEQ 30; 此外，還找到一個可能為AIP的基因SEQ 11。除了利用上述方法外，利用分子生物學的方式，如degenerate primer，可以在Lb. rhamnosus TCELL-1中發現AIP。所有找到的基因都利用類似pET-32a載體，pAB，表現，並直接處理和進行細菌素活性測試。這個實驗也使用His-taq 親和性管柱和陽離子交換樹酯去得到更純的胜肽，以避免干擾實驗的準確性。SEQ 11和SEQ 37接著還會進行autoinducing能力測試、生物膜形成。結果發現SEQ 34會抑制Listeria innocua ATCC 33091 、Citrobacter freundii ATCC 8090、Enterococcus faecalis ATCC 19433…等菌的生長; SEQ 11則是另外一個有活性並會殺死Listeria innocua ATCC 33091的胜肽，雖然它一開始被歸類成AIP。而就我們所知，過去鮮少有文獻也是利用搜尋基因的方式去找到新的細菌素。

論文指導教授推薦書------------------------------------------------------
學位考試委員會審定書---------------------------------------------------
英文摘要---------------------------------------------------------------------
中文摘要---------------------------------------------------------------------
誌謝辭------------------------------------------------------------------------
目錄---------------------------------------------------------------------------
前言---------------------------------------------------------------------------
材料與方法------------------------------------------------------------------
結果---------------------------------------------------------------------------
討論---------------------------------------------------------------------------
表1. 質體--------------------------------------------------------------------
表2. 基因名稱與分子量大小預測--------------------------------------
表3. 設計degenerate primer 05參考的基因序列---------------------
表4. SEQ 13 上下游基因-------------------------------------------------
表5. 粗略活性測試--------------------------------------------------------
表6. 以純化後的peptide進行較精確的活性測試 ------------------
表7. SEQ 11為AIP的證據-----------------------------------------------
圖1. Carnobacterium piscicola LV17B產生bactericoin的機制----
圖2. Degenerate primer對Lb. rhamnosus TCELL-1所做的PCR結果---------------------------------------------------------------------------
圖3. HPK、autoinducer、transporter在Lb. rhamnosus TCELL-1的相對位置------------------------------------------------------------------
圖4. 找尋Lb. rhamnosus TCELL-1上autoinducer基因-----------
圖5. pAB的製作過程------------------------------------------------------
圖6. 此為各個待表現peptide的DNA片段--------------------------
圖7. mSEQ 35基因的合成方法------------------------------------------
圖8. pUCB-HR的建構圖--------------------------------------------------
圖9. 各個目標蛋白的電泳圖--------------------------------------------
圖10. 各個純化的pepide經過thrombin處理後的電泳圖---------
圖11. 證實Lb. caseiVec確實含有pUCB-HR ------------------------
圖12. Lb. casei與Lb. caseiVec培養在50 μg/ml mSEQ 11的生長曲線------------------------------------------------------------------------
圖13. mSEQ 11對gusA活性的影響------------------------------------
圖14. mSEQ 11與mSEQ 37對部份菌株生成biofilm的影響-----
附錄1. 菌種及培養條件--------------------------------------------------
附錄2. 各種培養基和試劑-----------------------------------------------
附錄3. 此論文中所有使用過的primer --------------------------------
參考文獻---------------------------------------------------------------------

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