臺灣博碩士論文加值系統

在實驗室先前的研究發現在BJ2168菌株中過度表現ADH3p會使得粒線體型態產生分裂的狀況，但是BJ2168為蛋白酶缺陷菌株，為了驗證在一般菌株亦有一樣效果，本實驗使用BY4742和ΔADH3（BY4741）菌株過度表現ADH3p來驗證此結果，最後發現在BY4742菌株中亦有同樣結果，但是在ΔADH3菌株中，可能因為ADH3p表現量不足，導致粒線體型態呈現融合的狀態。

In previous studies, we found that overexpression ADH3p in BJ2168 will let mitochondria fission. BJ2168 is protease-deficient stain. In order to verify the result in BY4742 and ΔADH3(BY4741) strain, we did the same experiments to verify the results. Finally, in BY4742 strain, it had the same result. But in ADH3Δstrain, it looked different. It might be due to insufficient amount ADH3p.

目錄
謝誌I
中文摘要 II
英文摘要 III
圖目錄 VI
表目錄 VII
ADH3p的表現在酵母菌BY4742和ΔADH3（BY4741）中對粒線體型態的影響 1
第一章 緒論 1
1.1 前言 1
1.2 文獻回顧 2
1.3 研究動機 3
第二章 材料與方法 4
2.1 實驗材料 4
2.1.1 菌株 4
2.1.2 實驗藥品器材 4
2.1.3 實驗儀器 5
2.2 實驗方法 5
2.2.1 E. coli 質體抽取 6
2.2.2 S. cerevisiae 勝任細胞製備 6
2.2.3 S. cerevisiae transformation（heat shock method） 7
2.2.4 菌種保存 8
2.2.5 粒線體型態觀察 9
2.2.6 SDS-PAGE分析 10
第三章 實驗結果 11
3.1 粒線體型態觀察 11
3.2 SDS-PAGE分析 13
第四章 討論 16
附錄 17
一、 酵母菌隨機突變 17
二、 常用藥品組成 44
LB（Solid） 44
LB（Liquid） 44
Phosphate-buffered saline（PBS） 44
SC-dropout（Liquid） 45
dropout（Solid） 45
SDS running buffer 45
Separating gel（SDS-page下層膠） 46
Stacking gel（SDS-page上層膠） 46
YPD medium 46
YPD plate 47
三、 載體 47
1. pGEMR-T Easy Vector（Promega）[41] 47
參考資料 48

圖目錄
第一部分ADH3p的表現在酵母菌BY4742和ΔADH3（BY4741）中對粒線體型態的影響
圖1 粒線體氧化磷酸化示意圖 1
圖2 實驗流程 6
圖3 過度誘導ADH3p一天於BY4742的粒線體型態分裂比例 11
圖4 過度誘導ADH3p一天於BY4742的粒線體型態 12
圖5 過度誘導ADH3p三天於ΔADH3（BY4741）的粒線體型態分裂比例 12
圖6 過度誘導ADH3p 三天於ΔADH3（BY4741）的粒線體型態 13
圖7過度誘導ADH3p的SDS-page(菌株：BY4742、誘導時間:1天、電泳條件：120V 2.5小時、ADH3-GFP 分子量: 67.27 kDa) 14
圖8過度誘導ADH3p的SDS-page(菌株：ΔADH3(BY4741)、誘導時間:3天、電泳條件：200V 1小時、ADH3-GFP 分子量: 67.27 kDa) 15

表目錄
ADH3p的表現在酵母菌BY4742和ΔADH3(BY4741)中對粒線體型態的影響
表1 TE/LiOAc/DTT buffer配方 7
表2 PTLAD配方 8

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