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大腸菌全蛋白の細胞内局在性の網羅的な観察

全面观察所有大肠杆菌蛋白的细胞内定位

基本信息

批准号：
15013259
负责人：
小方康至
金额：
$ 1.98万
依托单位：
National Institute of Genetics
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2003
资助国家：
日本
起止时间：
2003 至无数据
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15013259/
关键词：
大腸菌 ORF 蛍光蛋白細胞内局在性 y-gene 核様体輝点 GFP融合蛋白

项目摘要

私達は、大腸菌(W3110株)の全ORF(4,388)と蛍光蛋白(GFP)遺伝子の融合遺伝子を発現させるプラスミドのライブラリー("Archive clone library")(奈良先端科学技術大学院大学・森浩禎博士より分与)を用いて、大腸菌全蛋白の細胞内局在を網羅的に観察する事に依り、(1)大腸菌全蛋白を細胞内局在性に基付いて分類し、(2)既に機能が知られている蛋白の細胞内局在性から、《蛋白の細胞内局在性と機能の相関》を見出し、更に、此の《相関》に基付いて、(3)今だ機能未知の遺伝子(所謂"y-gene")産物の細胞内局在性から、其の機能を推定する事を目指している。《蛋白の細胞内局在性と機能の相関》が見出された蛋白や《蛋白の機能と相関の有る細胞内局在性》を示す"y-gene"産物の内の46種類に就いて、短い相同配列間で起こる組換えを利用したWanner法を用い、夫々のstop codonをgfp遺伝子に置き換え、染色体上から発現されたGFP融合蛋白の細胞内局在を観察した処、其れ等の内の12種類の蛋白はプラスミド上から発現されたGFP融合蛋白の細胞内局在と一致しており、夫々の細胞内局在性は、D.細胞内全体(4/5)、F.核様体(1/4)、J.輝点(5/12)、I.膜と輝点(1/10)、L.核様体と輝点(1/1)であった。此処で、プラスミド上から発現されたGFP融合蛋白の細胞内局在性がF.核様体やJ.輝点であったにも拘わらず、染色体上から発現されたGFP融合蛋白の細胞内局在性が其れとは異なる蛋白の殆ど(夫々3/3と6/7)はD.細胞内全体に分類された。又、プラスミド上から発現されたGFP融合蛋白の細胞内局在性がK.リング(繊維状)であった蛋白に就いては、染色体上から発現されたGFP融合蛋白の細胞内局在性はK.リング(繊維状)では無く(0/7)、其れ等の半分はJ.輝点(3/7)に、残りの半分はD.細胞内全体(3/7)に分類された。従って、プラスミド上から発現されたGFP融合蛋白の細胞内局在性の内、染色体上から発現されたGFP融合蛋白の細胞内局在性と良く一致するのはD.細胞内全体であって、F.核様体やJ.輝点に分類された蛋白の中にはD.細胞内全体に存在する蛋白が有る可能性が残る。

The whole ORF (4388) phosphoprotein (GFP) protein (GFP) fusion protein of private and giant fungus (W3110 strain) was fused to detect the whole protein protein ("Archive clone library") (Mori Haoyi, University of Nara University of Advanced Science and Technology). The main results are as follows: (1) the whole protein of Magnoliae spp. (1) the whole protein of Magnoliae is classified into different classes. (2) the mechanism is able to know that the protein is not present in the cell, and that the protein in the cell is different from each other. (1) the whole protein of Magnoliae spp. (1) the whole protein of Magnoliae is classified into different classes. (2) it is possible to know that the protein is present in the cell. (2) it is possible to know that the protein is present in the cell. Its mechanism is capable of presuming that the incident is intended to refer to the accident. It can be seen that there is no protein in the cell. The protein can be detected in the cell. The protein can be detected in the y-gene. It is shown that there is no protein in the cell. In the same column, it is necessary to use the Wanner method to determine the concentration of the protein in the same column. By using the method of Wanner, it is possible to detect the protein in the cell by using the enzyme Wanner method. The GFP fusion protein was detected on the chromosome. The intracellular location of the GFP fusion protein was consistent with that of the other 12 proteins in the chromosome. The whole cell (4thumb 5), F. Nuclear body (1 stroke 4), J. Point (5 beat 12), I. Membrane temperature point (1 stroke 10), L. The nuclear body temperature point (1 ram 1) is closely related to the body temperature. The expression of GFP fusion protein was detected in this experiment. The nuclear body J. The expression of GFP fusion protein was detected on the chromosome, and the intracellular activity of the fusion protein was detected. All the members of the cell are classified and classified. In addition, it was found that the GFP fusion protein was present in the cells of K. The expression of GFP fusion protein was detected on chromosomes, and the intracellular locality of K. The system is free of charge (0 inch 7), and its equivalent is equal to half of the total value of J. Point (3 beat 7), stump half D. The whole body of cells (3ppm 7) was classified into different categories. The GFP fusion protein was detected on the chromosome and chromosome, and the GFP fusion protein was localized in the cell. The whole body of the cell, F. The nuclear body J. In the classification of protein, there is no difference between two groups. The existence of protein in the whole cell has the possibility of residual protein.