畢業論文

打賞
當前位置: 畢業論文 > 生物論文 >

水稻ABERRANT PANICLE ORGANIZATION 1 (APO1)/ ABERRANT PANICLE ORGANIZATION 2(APO2)互作基因

時間:2019-06-24 19:57來源:畢業論文
水稻ABERRANT PANICLE ORGANIZATION 1 (APO1)/ ABERRANT PANICLE ORGANIZATION 2(APO2)互作基因突變體的構建和功能分析

摘要:[目的]水稻是重要的糧食作物,水稻穗型與產量緊密相關。研究控制水稻穗型的基因功能以及發掘新的功能基因對提高水稻產量有重要意義。APO1與APO2蛋白可以直接相互作用并共同調控水稻幼穗分化。APO1是水稻的F-box蛋白,介導底物蛋白質的泛素化降解。本研究分析了APO1、APO2/RFL的功能,試圖明確APO1和APO2/RFL的互作蛋白,以及構建與APO1和APO2/RFL互作基因的突變體。[方法]利用已有的apo1和apo2/rfl突變體,觀察突變表型。并將mcherry蛋白基因與APO1連接,GFP蛋白基因與APO2/RFL連接,分別導入apo1和apo2/rfl突變體,觀察表型恢復狀況。提取apo1突變體和野生型水稻的幼穗蛋白,進行iTRAQ分析,獲得差異表達蛋白質。后進行酵母雙雜篩選,明確互作蛋白。利用CRISPR/Cas9技術構建互作基因突變體。[結果]與野生型水稻相比,apo1和apo2/rfl突變體的穗形明顯變小,二次枝梗數也明顯減小。apo2/rfl突變體導入gRFL:GFP以后,穗型出現一定程度的恢復。apo1突變體導入gAPO1:mchery后,APO1基因出現超表達,穗型較野生型明顯變大。iTRAQ分析發現apo1與野生型的幼穗有279個蛋白表達存在顯著差異,包括細胞分裂、分化與代謝等多方面的蛋白。酵母雙雜篩選得到了與APO1和APO2/RFL蛋白直接互作的蛋白質。CRISPR/Cas9技術構建了多個突變體。 [結論]明確了APO1和APO2/RFL基因在蛋白層面對水稻穗分化的影響,為下一步了解它們的互作機制奠定基礎。36602
關鍵詞:水稻;APO1;APO2/RFL;iTRAQ;酵母雙雜;CRISPR/Cas9
The construct of the mutants interact with ABERRANT PANICLE ORGANIZATION 1 (APO1)/ABERRANT PANICLE ORGANIZATION 2 (APO2/RFL)of rice and the analyze of the function
Abstract: [Objectives] rice is an important crop. The shape of spike has a tight connection to the production. With which, do research about the genes of rice spike and dig out new function gene so that improve the production has a good meaning. The proteins of APO1 and APO2/RFL could interact with each other and cooperate to control the dedifferentiation of young spike of rice. APO1 is F-box protein of rice, mediate substrate protein ubiquitination degradation. The research analyzes the function of APO1 and APO2/RFL, try to make it clear about the substrate protein which APO1 degradation. At the same time, construct the mutants of the genes interact with APO1 and APO2/RFL. [Methods] make use of apo1 and apo2/rfl mutants have gained. First, observe the phenotypes compared with wild type. And connect the gene of mchery protein with APO1,connect the gene of GFP with APO2/RFL,turned them into apo1 and apo2/rfl mutant respectively. Observe the phenotype whether it recovers. Then, extract the proteins of young spikes of apo1 mutant and wild type, analyze with the technology of iTRAQ. Then filter by the technology of yeast two hybrid, so that gain the proteins interact with each other. Then we use the technology of CRISPR/Cas9 to construct the mutants of the genes interact with APO1 and APO2/RFL. [Results] Compared with wild type, the shape of spike of apo1 mutant was obviously smaller, the amount of secondary branches was also less. When gRFL:GFP turned into apo2/rfl mutant, the phenotype of spike was recovered to a certain extent. When gAPO1:mchery turned into apo1 mutant, the APO1 gene was overexpressed, with the phenotype of spike become bigger. The analyze of iTRAQ found that there are 279 proteins express with obvious difference, including cell pision, cell dedifferentiation, and metabolism, and so on. We also did Yeast two hybrid, to find the proteins which gene APO1 and APO2/RFL controls directly. The technology of CRISPR/Cas9 has constructed some mutants. [Conclusions] We demonstrated how APO1 and APO2/RFL have an influence at rice spike dedifferentiation at the angle of protein, which makes a base on the research on how APO1 and APO2/RFL interaction.
源¥自%六^^維*論-文+網=www.aftnzs.live

Keywords: rice; APO1; APO2/RFL; iTRAQ; Yeast two hybrid;CRISPR/Cas9 水稻ABERRANT PANICLE ORGANIZATION 1 (APO1)/ ABERRANT PANICLE ORGANIZATION 2(APO2)互作基因:http://www.aftnzs.live/shengwu/20190624/35121.html
------分隔線----------------------------
推薦內容
双色球走势图带连线 新疆11选5前三走势图 体育彩票环岛赛玩法 老版吉祥棋牌下载安装 上海快3智能号码推荐 湖北快三奖结果查询 上海时时彩信誉平台 黑龙江p62走势透图 湖南幸运赛车 河南麻将群怎么进入 北京pk赛车官方平台 德甲积分榜上赛季 浙江快乐彩开奖直播 单机版麻将游戏 天星山西麻将苹果授权码 秒速快三是骗局吗 江苏十一选五遗漏图