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高等植物におけるタンパク質分泌の分子機構-条件優性体細胞遺伝学によるアプローチ

高等植物蛋白质分泌的分子机制——一种利用条件显性体细胞遗传学的方法

基本信息

批准号：
08874109
负责人：
中野明彦
金额：
$ 0.32万
依托单位：
The Institute of Physical and Chemical Research (1997)The University of Tokyo (1996)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Exploratory Research
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08874109/
关键词：
タンパク質の分泌小胞輸送 SAR1 RMA1 GDI 優性致死変異タバコシロイヌナズナ ARA4

项目摘要

植物細胞において,タンパク質の分泌過程は,細胞骨格系と協同して細胞の極性と伸長方向を決定していくという重要な意義を持つ,本研究では,植物における分泌の研究に,酵母で大きな成功を見ている分子遺伝学的アプローチを導入することを目指した.タバコとシロイヌナズナのcDNAを酵母で発現する実験系を構築し,以下の研究を行った.1,小胞体-ゴルジ体間輸送に働く酵母遺伝子SAR1のタバコとシロイヌナズナの相同遺伝子を得,これが酵母SAR1欠失株を相補することを利用してさまざまな変異の解析を行った.その結果,H74LとN129Iという変異アレルが植物の野生型SAR1に対して優性致死に働くことを明らかにし,植物で発現調節することにより条件致死変異株を作成する展望を開いた.2,GAL1プロモーターの支配下に酵母内で条件的に発現するシロイヌナズナcDNAライブラリーを構築し,これを用いて酵母sec15ts変異を抑圧するシロイヌナズナ遺伝子RMA1を同定した.この遺伝子は,RING fingerモチーフを持つ新しいタイプの膜タンパク質をコードしていた.3,シロイヌナズナの低分子量GTPase Ara4を発現することが酵母のypt1,ypt3,sec4変異株の生育を悪くすることを利用して得た,Ara4の制御因子AtGDIについて,さらに解析を進め,クロスハイブリダイゼーションによって第2の遺伝子AtGDI2を同定した.

The process of cell secretion is very difficult in plants, and the cellular bone lattice works in conjunction with the direction of cell growth to determine the important meaning of cell growth. In this study, the secretion of plant cells was studied, and the yeast was successfully identified by the molecular biology of yeast. In the following study, the following studies were carried out. 1. The small-cell body-to-body SAR1 of yeast was transferred to the body of yeast, and the mutant strain of yeast SAR1 was used to obtain the same result, and the mutant strain of yeast SAR1 was analyzed by scanning electron microscopy (RIA). The results showed that H74LNAs N129IRNA isolated from plants showed that wild-type SAR1 was found to be sexually lethal. The results showed that the plants showed that they were killed under the condition of conditional death. The results showed that the plants were divided into two groups: the wild type, the wild type, the wild type, The yeast sec15ts was used to control the growth of the yeast. The RMA1 of the yeast was determined by the same method. The main results are as follows: (1) the growth of yeast ypt1,ypt3,sec4 strain was detected by low molecular weight GTPase Ara4 (low molecular weight). The results showed that the growth rate of yeast strain was significantly higher than that of yeast strain (ypt1,ypt3,sec4 strain). The results showed that the growth rate of yeast strain was significantly higher than that of low molecular weight GTPase Ara4 (low molecular weight). The results showed that the growth rate of yeast strain was significantly higher than that of yeast strain (Saccharomyces cerevisiae). The results showed that Ara4 control factor AtGDI was successfully used, and the results were analyzed. I don't know what to say. I don't know what to do. I don't know what to do with the AtGDI2.