請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62014
標題: | 綠竹 BoMSP41 之細胞內定位與生化性質探討 Studies on the subcellular localization and biochemical properties of bamboo BoMSP41 |
作者: | Fang-Chieh Yang 楊芳潔 |
指導教授: | 王愛玉(Ai-Yu Wang) |
共同指導教授: | 洪傳揚(Chwan-Yang Hong) |
關鍵字: | 綠竹,單子葉植物特有蛋白質,細胞內定位,葉綠體,內生性不具穩定構形蛋白質, bamboo,monocot-specific protein,subcellular localization,chloroplast,intrinsic disorder protein, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | BoMSP41 (Monocot-specific protein-41 in Bambusa oldhamii) 為綠竹之未知功能基因,於綠竹快速生長時期,其基因表現會大幅提升。BoMSP41 胺基酸組成富含 Gln, Glu, Lys, Arg 及 Asp,且含有一些重複的片段。此蛋白質被預測為一個內生性不具穩定構形的蛋白質 intrinsic disorder protein (IDP) 。為了瞭解 BoMSP41 於細胞內存在的位置,本研究建構了以玉米 Ubiquitin 啟動子驅動表現的 GFP::BoMSP41 的重組質體,並對水稻進行轉殖,使 GFP::BoMSP41 融合蛋白質恆常表現於轉殖水稻中。以共軛焦顯微鏡觀察轉殖水稻的原生質體,結果顯示 GFP::BoMSP41 可能被運送至成熟的葉綠體中,推測 BoMSP41 可能參與光合作用、質體基因表現或在葉綠體中發生的其他代謝路徑。為了探討 BoMSP41 的生化性質,以大腸桿菌表現帶有 His-tag 的BoMSP41。由 SDS-聚丙烯醯胺膠體及西方點墨法結果顯示: BoMSP41 於SDS 膠體上的泳動率比預期的分子量應有的泳動率低。此外, BoMSP41 可被 3% TCA沉澱下,但對熱具較高耐受性。泳動率和熱穩定性高這兩點特性與許多 IDP 特性相符。然而,其結構及生化性質仍需進一步探討。 BoMSP41 (Monocot-specific protein-41 in Bambusa oldhamii) is an unknown function gene highly up-regulated in rapidly growing bamboo shoots. The BoMSP41-encoding protein contains tandemly repeated sequences and a high proportion of Gln, Glu, Lys, Arg, and Asp. It was predicted to be an intrinsic disorder protein (IDP). To investigate the localization of BoMSP41 in cells, transgenic rice plants constitutively expressing the GFP::BoMSP41 fusion protein under the control of a maize ubiquitin promoter were constructed. Analysis of protoplasts isolated from seedlings of the transgenic plants by confocal laser scanning microscopy indicated that the GFP::BoMSP41 fusion proteins were transported into mature chloroplasts, suggesting that BoMSP41 may participate in photosynthesis, plastid gene expression or other metabolic pathways occurred in chloroplasts. To characterize the biochemical properties of BoMSP41, the His-tagged BoMSP41 recombinant protein was expressed in Escherichia coli. SDS-PAGE and western analyses showed that the electrophoretic mobility of the recombinant BoMSP41 protein was lower than expected. In addition, the recombinant BoMSP41 protein could be precipitated by 3% TCA but exhibited high thermostability. The lower electrophoretic mobility and high thermostability of BoMSP41 are in agreement with the general properties of many IDPs; however, the structure and the biochemical identity of this protein require further investigation. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62014 |
全文授權: | 有償授權 |
顯示於系所單位: | 生化科技學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 2.16 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。