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大豆疫霉效應分子Avh241的寄主靶標鑒定

時間:2019-04-16 21:05來源:畢業論文
探究了大豆疫霉效應分子Avh241的功能,通過篩選其寄主靶標,并從中選擇候選互作蛋白,利用Co-IP和GST Pull-Down兩種技術對其進行了互作驗證,成功篩選到Avh241的互作蛋白GmHin1

摘要:大豆疫霉引起的大豆根莖腐病是大豆的毀滅性病害之一,嚴重威脅全世界的大豆生產。在侵染植物的過程中,大豆疫霉會分泌大量的效應分子進入寄主體內干擾植物免疫,促進自身侵染。因此,深入探究效應分子干擾植物抗病性的機理,明確大豆疫霉致病的途徑以及作用在寄主植物的靶標位點,可以為有效的病害防控策略提供新的思路及方向,同時對設計和改造抗病性有重要意義。本文探究了大豆疫霉效應分子Avh241的功能,通過篩選其寄主靶標,并從中選擇候選互作蛋白,利用Co-IP和GST Pull-Down兩種技術對其進行了互作驗證,成功篩選到Avh241的互作蛋白GmHin1。該研究為進一步理解植物與病原菌的互作機制提供了理論依據。34617
畢業論文關鍵詞:效應分子;Avh241;寄主靶標;蛋白互作;Co-IP;GST Pull-Down
Identification of Host target of Phytophthora sojae effector Avh241
Abstract: Phytophthora sojae causes the Phytophthora root and stem rot disease of soybean. P. sojae delivers varieties of effectors into host cell to reprogram host immunity during infecting host plant. Therefore, it’s significant to learn the mechanism of how effectors interfere plant immunity and the way that P. sojae infect the host plant, which could provide guidance about control methods and strategies of plant diseases. Avh241 is one of the RxLR effectors from P. sojae, which can promote P. sojae infection on its host plant. Our research studies analyzes the function of Avh241. In this study, we screen out the host target protein which may interact with Avh241 and select one candidate interacting protein of Avh241 to do the research. We also provide the interaction between Avh241 and candidate interacting protein by Co-IP and GST Pull-Down. Our result is that we find an interacting protein named hin1 and demonstrate that it can interact with Avh241. 源¥自%六^^維*論-文+網=www.aftnzs.live
Key words: effector;Avh241;Host target;Interacting protein;Co-IP;GST Pull-Down
目  錄
摘要    1
關鍵詞    1
Abstract    1
Key words    1
引言    1
材料與方法    3
1.1  供試植物和供試菌株的保存和培養    3
1.2  目標基因的克隆及重組質粒的構建    3
1.2.1 目的基因的克隆    3
1.2.2  重組質粒的構建    3
1.3  大腸桿菌JM109感受態細胞的制備和轉化    4
1.3.1  大腸桿菌JM109感受態細胞的制備    4
1.3.2  熱擊轉化大腸桿菌JM109感受態細胞    4
1.4  大腸桿菌質粒提取    5
1.5  農桿菌GV3101感受態細胞的制備和轉化    5
1.5.1  農桿菌GV3101感受態細胞的制備    5
1.5.2  電擊轉化農桿菌GV3101感受態細胞    5
1.6  目標基因的植物表達    6
1.6.1  注射煙草用緩沖液的配制(50 mL)    6
1.6.2  注射煙草用緩沖液懸浮農桿菌    6
1.6.3  注射煙草    6
1.7  植物蛋白的提取    6
1.8  蛋白樣品Western 驗證    7
1.9 蛋白互作驗證    7
1.9.1 植物體內免疫共沉淀技術(Co-IP)    7
1.9.2  蛋白質Pull-Down技術    8
2  結果與分析    9
2.1  重組質粒pBinGFP-Avh241的構建    9
2.2  候選互作蛋白篩選結果    10 源¥自%六^^維*論-文+網=www.aftnzs.live
2.3  Avh241候選互作蛋白的克隆及重組質粒的構建    11 大豆疫霉效應分子Avh241的寄主靶標鑒定:http://www.aftnzs.live/shengwu/20190416/32179.html
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