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標題: 以生物技術改良及創新觀賞花卉百合之花形
Generation of Flowers with Novel Phenotypes in Lily by Using Methods of Biotechnology
作者: 楊長賢
關鍵字: 生物技術, 園藝;應用研究
In this project, constructs contained various forms of LAG cDNA will be generated and transformed into tobacco and Eustoma grandiflorum for further functional analysis.The phenotypes observed in transgenic plants will be examined to see if any floral organ conversion such as sepal to carpel or petal to stamen were generated.Any dominant negative mutations produced by transforming truncated LAG will also be analyzed.The results will help to confirm the function for LAG.Furthermore, any interested novel flower phenotype observed in transgenic E.grandiflorum will be evaluated for the potential use in the flower market directly.In addition, LAG proteins will be purified from bacterial expression system and used to generate antibody for further Western blot analysis.In this project, we also plan to clone the promoter region for LAG gene using IPCR strategy.The cloned promoter will be fused to report gene such as GUS or GFP and transformed into plants for expression analysis.The constructs described above will be transformed into lily using genegun approach to initiate the functional analysis for LAG in lily.

本年度之工作目標將構築完成含LAG基因cDNA之各式構築體, 透過膿桿菌轉殖入菸草及洋桔梗中.並分析轉殖菸草及洋桔梗中LAG基因表現之情形, 及其整體性狀, 觀察是否花器中花萼及花瓣有轉形至雌蕊及雄蕊之形態.另外也將觀察轉含不同缺失之構築體是否會有dominant negative mutantion的出現, 以便說明在阿拉伯芥中得到的結果是否正確及具代表性.所得到的轉基因洋桔梗植株, 可用在時實際市場推廣到用.在LAG基因之表現探討上, 將進一步在細菌中表現及純化LAG之蛋白質, 並製備LAG之抗體, 透過Western blot之方法分析LAG之蛋白質表現與其mRNA之表現之相關性.本年度並將透過IPCR ( inverse PCR )的方式選殖出LAG基因之調控區( promoter ), 選殖出之LAG promoter將來可用來在花器中專一表達任何外來之基因產物, 在植物生產上有極高的應用價值以供將來分析應用.另外我們亦將進行LAG在鐵砲百合中轉殖之工作.我們計畫在本年度中將含LAG的各式構築體透過基因槍的方法先進行一些轉殖測試工作.
其他識別: 90農科-2.1.1-糧-Z1(10)
Appears in Collections:生物科技學研究所

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